LOCUS       40924_6182        7507 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Single-copy E. coli plasmid vector designed for constructing 
            Bacterial Artificial Chromosomes (BACs).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7507)
  AUTHORS   New England Biolabs
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7507)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     289..305
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        312..330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    333..389
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(396..414)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(432..448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(456..472)
                     /label=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)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(525..546)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(769..1425)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1426..1528)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      2455..2674
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             2765..3517
                     /codon_start=1
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    3523..3773
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             4099..5271
                     /codon_start=1
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             5274..6242
                     /codon_start=1
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    6318..6791
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    complement(7051..7449)
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(7467..7500)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
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
//
