pBeloBAC11

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  • Model: PVT10574
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pBeloBAC11

PVT10574    2ug

 

pBeloBAC11  Information

Replicon: OriS

Plasmid size: 7507bp

Prokaryotic resistance: chloramphenicol Chl

Clone strain: Escherichia coli DH5 alpha

Culture conditions: 37 C, aerobic, LB

Remarks: 1.bacterial artificial chromosome BAC plasmid
                2. Plasmid Maxi Preparation

 

pBeloBAC11 Description

Bacterial artificial chromosome,BAC is a kind of bacterial chromosome cloning vector constructed on the basis of F-plasmid, named pBACs. Its capacity is between 50-300 kb. All kinds of pBACs are single-copy in E. coli.

BAC is a synthetic carrier based on the F factor of Escherichia coli. Factor F, also known as plasmid F, is a "plasmid" that transfers natural host genes to another host cell. F factor replication in E. coli is strictly controlled and maintained at a low copy, usually 1-2 copies per cell. Its parB gene can reject the exogenous F plasmid in the cell, restrict the rearrangement of plasmids between molecules, and reduce the level of drainage and chimerism of exogenous DNA in the recombinant BAC. In addition, F factor has the potential to carry 1 Mbp insert fragment, which makes it possible to construct a large-capacity cloning BAC vector.

BAC carriers mainly include oriS, repE (control F plasmid replication) and parA, parB (control copy number) and other components. BAC-based vectors have been widely accepted by the scientific community for their low frequency of chimerism, high conversion efficiency, and circular structure in bacteria, which is easy to isolate and purify. At present, it is mainly used in the construction of large fragment genomic library and the study of large gene cluster, and plays an important role in various biological genome projects.

 

pBeloBAC11 Sequence

LOCUS       Exported                7507 bp ds-DNA     circular SYN 19-MAY-2017

DEFINITION  Single-copy E. coli plasmid vector designed for constructing 

            Bacterial Artificial Chromosomes (BACs).

ACCESSION   U51113

VERSION     .

KEYWORDS    pBeloBAC11

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 7507)

  AUTHORS   New England Biolabs

  TITLE     Direct Submission

FEATURES             Location/Qualifiers

     source          1..7507

                     /organism="synthetic DNA construct"

                     /lab_host="Escherichia coli"

                     /mol_type="other DNA"

     CDS             complement(56..436)

                     /codon_start=1

                     /gene="lacZ (fragment)"

                     /product="LacZ-alpha fragment of beta-galactosidase"

                     /note="lacZ-alpha"

                     /translation="MTMITPSYLGDTIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL

                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLL

                     THLCGISHRIWCTLSTICSDAA"

     primer_bind     289..305

                     /note="M13 fwd"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     promoter        312..330

                     /note="T7 promoter"

                     /note="promoter for bacteriophage T7 RNA polymerase"

     misc_feature    333..389

                     /note="MCS"

                     /note="pUC18/19 multiple cloning site"

     promoter        complement(396..414)

                     /note="SP6 promoter"

                     /note="promoter for bacteriophage SP6 RNA polymerase"

     primer_bind     complement(432..448)

                     /note="M13 rev"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     protein_bind    456..472

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="lac operator"

                     /note="The lac repressor binds to the lac operator to 

                     inhibit transcription in E. coli. This inhibition can be 

                     relieved by adding lactose or 

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

     promoter        complement(480..510)

                     /note="lac promoter"

                     /note="promoter for the E. coli lac operon"

     CDS             complement(766..1425)

                     /codon_start=1

                     /gene="cat"

                     /product="chloramphenicol acetyltransferase"

                     /note="CmR"

                     /note="confers resistance to chloramphenicol"

                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL

                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS

                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM

                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"

     promoter        complement(1426..1528)

                     /note="cat promoter"

                     /note="promoter of the E. coli cat gene"

     rep_origin      2455..2674

                     /note="ori2"

                     /note="secondary origin of replication for the bacterial F 

                     plasmid; also known as oriS"

     CDS             2765..3520

                     /codon_start=1

                     /gene="repE"

                     /product="replication initiation protein for the bacterial 

                     F plasmid"

                     /note="repE"

                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ

                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG

                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM

                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM

                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"

     misc_feature    3523..3773

                     /gene="incC"

                     /note="incC"

                     /note="incompatibility region of the bacterial F plasmid"

     CDS             4099..5274

                     /codon_start=1

                     /gene="sopA"

                     /product="partitioning protein for the bacterial F plasmid"

                     /note="sopA"

                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI

                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA

                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL

                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD

                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF

                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE

                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"

     CDS             5274..6245

                     /codon_start=1

                     /gene="sopB"

                     /product="partitioning protein for the bacterial F plasmid"

                     /note="sopB"

                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA

                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP

                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER

                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD

                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF

                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"

     misc_feature    6318..6791

                     /gene="sopC"

                     /note="sopC"

                     /note="centromere-like partitioning region of the bacterial

                     F plasmid"

     misc_feature    7051..7449

                     /note="cos"

                     /note="lambda cos site; allows packaging into phage lambda 

                     particles"

     protein_bind    7467..7500

                     /bound_moiety="Cre recombinase"

                     /note="loxP"

                     /note="Cre-mediated recombination occurs in the 8-bp core 

                     sequence (GCATACAT)."

ORIGIN

        1 gcggccgcaa ggggttcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg

       61 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat

      121 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg

      181 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg

      241 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg

      301 ccagtgaatt gtaatacgac tcactatagg gcgaattcga gctcggtacc cggggatcct

      361 ctagagtcga cctgcaggca tgcaagcttg agtattctat agtgtcacct aaatagcttg

      421 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac

      481 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc

      541 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg

      601 cattaatgaa tcggccaacg cgaacccctt gcggccgccc gggccgtcga ccaattctca

      661 tgtttgacag cttatcatcg aatttctgcc attcatccgc ttattatcac ttattcaggc

      721 gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg

      781 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca

      841 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata

      901 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa

      961 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt

     1021 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa

     1081 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg

     1141 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc

     1201 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata

     1261 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt

     1321 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca

     1381 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc

     1441 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg

     1501 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg

     1561 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac

     1621 aggtatttat tcgcgataag ctcatggagc ggcgtaaccg tcgcacagga aggacagaga

     1681 aagcgcggat ctgggaagtg acggacagaa cggtcaggac ctggattggg gaggcggttg

     1741 ccgccgctgc tgctgacggt gtgacgttct ctgttccggt cacaccacat acgttccgcc

     1801 attcctatgc gatgcacatg ctgtatgccg gtataccgct gaaagttctg caaagcctga

     1861 tgggacataa gtccatcagt tcaacggaag tctacacgaa ggtttttgcg ctggatgtgg

     1921 ctgcccggca ccgggtgcag tttgcgatgc cggagtctga tgcggttgcg atgctgaaac

     1981 aattatcctg agaataaatg ccttggcctt tatatggaaa tgtggaactg agtggatatg

     2041 ctgtttttgt ctgttaaaca gagaagctgg ctgttatcca ctgagaagcg aacgaaacag

     2101 tcgggaaaat ctcccattat cgtagagatc cgcattatta atctcaggag cctgtgtagc

     2161 gtttatagga agtagtgttc tgtcatgatg cctgcaagcg gtaacgaaaa cgatttgaat

     2221 atgccttcag gaacaataga aatcttcgtg cggtgttacg ttgaagtgga gcggattatg

     2281 tcagcaatgg acagaacaac ctaatgaaca cagaaccatg atgtggtctg tccttttaca

     2341 gccagtagtg ctcgccgcag tcgagcgaca gggcgaagcc ctcgagtgag cgaggaagca

     2401 ccagggaaca gcacttatat attctgctta cacacgatgc ctgaaaaaac ttcccttggg

     2461 gttatccact tatccacggg gatattttta taattatttt ttttatagtt tttagatctt

     2521 cttttttaga gcgccttgta ggcctttatc catgctggtt ctagagaagg tgttgtgaca

     2581 aattgccctt tcagtgtgac aaatcaccct caaatgacag tcctgtctgt gacaaattgc

     2641 ccttaaccct gtgacaaatt gccctcagaa gaagctgttt tttcacaaag ttatccctgc

     2701 ttattgactc ttttttattt agtgtgacaa tctaaaaact tgtcacactt cacatggatc

     2761 tgtcatggcg gaaacagcgg ttatcaatca caagaaacgt aaaaatagcc cgcgaatcgt

     2821 ccagtcaaac gacctcactg aggcggcata tagtctctcc cgggatcaaa aacgtatgct

     2881 gtatctgttc gttgaccaga tcagaaaatc tgatggcacc ctacaggaac atgacggtat

     2941 ctgcgagatc catgttgcta aatatgctga aatattcgga ttgacctctg cggaagccag

     3001 taaggatata cggcaggcat tgaagagttt cgcggggaag gaagtggttt tttatcgccc

     3061 tgaagaggat gccggcgatg aaaaaggcta tgaatctttt ccttggttta tcaaacgtgc

     3121 gcacagtcca tccagagggc tttacagtgt acatatcaac ccatatctca ttcccttctt

     3181 tatcgggtta cagaaccggt ttacgcagtt tcggcttagt gaaacaaaag aaatcaccaa

     3241 tccgtatgcc atgcgtttat acgaatccct gtgtcagtat cgtaagccgg atggctcagg

     3301 catcgtctct ctgaaaatcg actggatcat agagcgttac cagctgcctc aaagttacca

     3361 gcgtatgcct gacttccgcc gccgcttcct gcaggtctgt gttaatgaga tcaacagcag

     3421 aactccaatg cgcctctcat acattgagaa aaagaaaggc cgccagacga ctcatatcgt

     3481 attttccttc cgcgatatca cttccatgac gacaggatag tctgagggtt atctgtcaca

     3541 gatttgaggg tggttcgtca catttgttct gacctactga gggtaatttg tcacagtttt

     3601 gctgtttcct tcagcctgca tggattttct catacttttt gaactgtaat ttttaaggaa

     3661 gccaaatttg agggcagttt gtcacagttg atttccttct ctttcccttc gtcatgtgac

     3721 ctgatatcgg gggttagttc gtcatcattg atgagggttg attatcacag tttattactc

     3781 tgaattggct atccgcgtgt gtacctctac ctggagtttt tcccacggtg gatatttctt

     3841 cttgcgctga gcgtaagagc tatctgacag aacagttctt ctttgcttcc tcgccagttc

     3901 gctcgctatg ctcggttaca cggctgcggc gagcgctagt gataataagt gactgaggta

     3961 tgtgctcttc ttatctcctt ttgtagtgtt gctcttattt taaacaactt tgcggttttt

     4021 tgatgacttt gcgattttgt tgttgctttg cagtaaattg caagatttaa taaaaaaacg

     4081 caaagcaatg attaaaggat gttcagaatg aaactcatgg aaacacttaa ccagtgcata

     4141 aacgctggtc atgaaatgac gaaggctatc gccattgcac agtttaatga tgacagcccg

     4201 gaagcgagga aaataacccg gcgctggaga ataggtgaag cagcggattt agttggggtt

     4261 tcttctcagg ctatcagaga tgccgagaaa gcagggcgac taccgcaccc ggatatggaa

     4321 attcgaggac gggttgagca acgtgttggt tatacaattg aacaaattaa tcatatgcgt

     4381 gatgtgtttg gtacgcgatt gcgacgtgct gaagacgtat ttccaccggt gatcggggtt

     4441 gctgcccata aaggtggcgt ttacaaaacc tcagtttctg ttcatcttgc tcaggatctg

     4501 gctctgaagg ggctacgtgt tttgctcgtg gaaggtaacg acccccaggg aacagcctca

     4561 atgtatcacg gatgggtacc agatcttcat attcatgcag aagacactct cctgcctttc

     4621 tatcttgggg aaaaggacga tgtcacttat gcaataaagc ccacttgctg gccggggctt

     4681 gacattattc cttcctgtct ggctctgcac cgtattgaaa ctgagttaat gggcaaattt

     4741 gatgaaggta aactgcccac cgatccacac ctgatgctcc gactggccat tgaaactgtt

     4801 gctcatgact atgatgtcat agttattgac agcgcgccta acctgggtat cggcacgatt

     4861 aatgtcgtat gtgctgctga tgtgctgatt gttcccacgc ctgctgagtt gtttgactac

     4921 acctccgcac tgcagttttt cgatatgctt cgtgatctgc tcaagaacgt tgatcttaaa

     4981 gggttcgagc ctgatgtacg tattttgctt accaaataca gcaatagtaa tggctctcag

     5041 tccccgtgga tggaggagca aattcgggat gcctggggaa gcatggttct aaaaaatgtt

     5101 gtacgtgaaa cggatgaagt tggtaaaggt cagatccgga tgagaactgt ttttgaacag

     5161 gccattgatc aacgctcttc aactggtgcc tggagaaatg ctctttctat ttgggaacct

     5221 gtctgcaatg aaattttcga tcgtctgatt aaaccacgct gggagattag ataatgaagc

     5281 gtgcgcctgt tattccaaaa catacgctca atactcaacc ggttgaagat acttcgttat

     5341 cgacaccagc tgccccgatg gtggattcgt taattgcgcg cgtaggagta atggctcgcg

     5401 gtaatgccat tactttgcct gtatgtggtc gggatgtgaa gtttactctt gaagtgctcc

     5461 ggggtgatag tgttgagaag acctctcggg tatggtcagg taatgaacgt gaccaggagc

     5521 tgcttactga ggacgcactg gatgatctca tcccttcttt tctactgact ggtcaacaga

     5581 caccggcgtt cggtcgaaga gtatctggtg tcatagaaat tgccgatggg agtcgccgtc

     5641 gtaaagctgc tgcacttacc gaaagtgatt atcgtgttct ggttggcgag ctggatgatg

     5701 agcagatggc tgcattatcc agattgggta acgattatcg cccaacaagt gcttatgaac

     5761 gtggtcagcg ttatgcaagc cgattgcaga atgaatttgc tggaaatatt tctgcgctgg

     5821 ctgatgcgga aaatatttca cgtaagatta ttacccgctg tatcaacacc gccaaattgc

     5881 ctaaatcagt tgttgctctt ttttctcacc ccggtgaact atctgcccgg tcaggtgatg

     5941 cacttcaaaa agcctttaca gataaagagg aattacttaa gcagcaggca tctaaccttc

     6001 atgagcagaa aaaagctggg gtgatatttg aagctgaaga agttatcact cttttaactt

     6061 ctgtgcttaa aacgtcatct gcatcaagaa ctagtttaag ctcacgacat cagtttgctc

     6121 ctggagcgac agtattgtat aagggcgata aaatggtgct taacctggac aggtctcgtg

     6181 ttccaactga gtgtatagag aaaattgagg ccattcttaa ggaacttgaa aagccagcac

     6241 cctgatgcga ccacgtttta gtctacgttt atctgtcttt acttaatgtc ctttgttaca

     6301 ggccagaaag cataactggc ctgaatattc tctctgggcc cactgttcca cttgtatcgt

     6361 cggtctgata atcagactgg gaccacggtc ccactcgtat cgtcggtctg attattagtc

     6421 tgggaccacg gtcccactcg tatcgtcggt ctgattatta gtctgggacc acggtcccac

     6481 tcgtatcgtc ggtctgataa tcagactggg accacggtcc cactcgtatc gtcggtctga

     6541 ttattagtct gggaccatgg tcccactcgt atcgtcggtc tgattattag tctgggacca

     6601 cggtcccact cgtatcgtcg gtctgattat tagtctggaa ccacggtccc actcgtatcg

     6661 tcggtctgat tattagtctg ggaccacggt cccactcgta tcgtcggtct gattattagt

     6721 ctgggaccac gatcccactc gtgttgtcgg tctgattatc ggtctgggac cacggtccca

     6781 cttgtattgt cgatcagact atcagcgtga gactacgatt ccatcaatgc ctgtcaaggg

     6841 caagtattga catgtcgtcg taacctgtag aacggagtaa cctcggtgtg cggttgtatg

     6901 cctgctgtgg attgctgctg tgtcctgctt atccacaaca ttttgcgcac ggttatgtgg

     6961 acaaaatacc tggttaccca ggccgtgccg gcacgttaac cgggctgcat ccgatgcaag

     7021 tgtgtcgctg tcgacgagct cgcgagctcg gacatgaggt tgccccgtat tcagtgtcgc

     7081 tgatttgtat tgtctgaagt tgtttttacg ttaagttgat gcagatcaat taatacgata

     7141 cctgcgtcat aattgattat ttgacgtggt ttgatggcct ccacgcacgt tgtgatatgt

     7201 agatgataat cattatcact ttacgggtcc tttccggtga tccgacaggt tacggggcgg

     7261 cgacctcgcg ggttttcgct atttatgaaa attttccggt ttaaggcgtt tccgttcttc

     7321 ttcgtcataa cttaatgttt ttatttaaaa taccctctga aaagaaagga aacgacaggt

     7381 gctgaaagcg agctttttgg cctctgtcgt ttcctttctc tgtttttgtc cgtggaatga

     7441 acaatggaag tccgagctca tcgctaataa cttcgtatag catacattat acgaagttat

     7501 attcgat

//
 

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