LOCUS       40924_40722        7648 bp DNA     circular SYN 01-JAN-1980
DEFINITION  BAC vector with inducible copy number, suitable for cloning toxic 
            inserts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7648)
  AUTHORS   Lucigen
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7648)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     This sequence does not include a 3.3-kb lacZ-sacB stuffer fragment 
            inserted at the origin.
            Linearization with an enzyme such as BamHI, HindIII, or EcoRI 
            removes the stuffer.
FEATURES             Location/Qualifiers
     source          1..7648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(61..79)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(105..148)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        151..253
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             254..910
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      complement(927..958)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     protein_bind    984..1017
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1035..1433
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     misc_feature    complement(1694..2167)
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(2243..3211)
                     /codon_start=1
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(3214..4386)
                     /codon_start=1
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    complement(4712..4962)
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(4968..5720)
                     /codon_start=1
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      complement(5811..6030)
                     /direction=LEFT
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      complement(6106..6720)
                     /direction=LEFT
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     terminator      complement(7487..7514)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        7573..7591
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 ccaccacagt gggatccgaa gcttggaatt cacgtgactt gaagtcgcgg ccgcactgac
       61 cctatagtga gtcgtattaa tttaaatcat accaacatgg tcaaataaaa cgaaaggctc
      121 agtcgaaaga ctgggccttt cgttttaatc tgatcggcac gtaagaggtt ccaactttca
      181 ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc
      241 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
      301 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
      361 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
      421 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc
      481 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
      541 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
      601 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
      661 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
      721 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
      781 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
      841 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
      901 gggcggggcg taacctaggt gacagaagtc aaaagcctcc ggtcggaggc ttttgacttt
      961 ctgctagatc ggccgcatcg aatataactt cgtataatgt atgctatacg aagttattag
     1021 cgatgagctc ggacttccat tgttcattcc acggacaaaa acagagaaag gaaacgacag
     1081 aggccaaaaa gctcgctttc agcacctgtc gtttcctttc ttttcagagg gtattttaaa
     1141 taaaaacatt aagttatgac gaagaagaac ggaaacgcct taaaccggaa aattttcata
     1201 aatagcgaaa acccgcgagg tcgccgcccc gtaacctgtc ggatcaccgg aaaggacccg
     1261 taaagtgata atgattatca tctacatatc acaacgtgcg tggaggccat caaaccacgt
     1321 caaataatca attatgacgc aggtatcgta ttaattgatc tgcatcaact taacgtaaaa
     1381 gcaacttcag acaatacaaa tcagcgacac tgaatacggg gcaacctcat gtccgagctc
     1441 gcgagctcgt cgacagcgac acacttgcat cggatgcagc ccggttaacg tgccggcacg
     1501 gcctgggtaa ccaggtattt tgtccacata accgtgcgca aaatgttgtg gataagcagg
     1561 acacagcagc aatccacagc aggcatacaa ccgcacaccg aggttactcc gttctacagg
     1621 ttacgacgac catgtcaata cttgcccttg acaggcattg atggaatcgt agtctcacgc
     1681 tgatagtctg atcgacaata caagtgggac cgtggtccca gaccgataat cagaccgaca
     1741 acacgagtgg gatcgtggtc ccagactaat aatcagaccg acgatacgag tgggaccgtg
     1801 gtcccagact aataatcaga ccgacgatac gagtgggacc gtggttccag actaataatc
     1861 agaccgacga tacgagtggg accgtggtcc cagactaata atcagaccga cgatacgagt
     1921 gggaccatgg tcccagacta ataatcagac cgacgatacg agtgggaccg tggtcccagt
     1981 ctgattatca gaccgacgat acgagtggga ccgtggtccc agactaataa tcagaccgac
     2041 gatacgagtg ggaccgtggt cccagactaa taatcagacc gacgatacga gtgggaccgt
     2101 ggtcccagtc tgattatcag accgacgata caagtggaac agtgggccca gagagaatat
     2161 tcaggccagt tatgctttct ggcctgtaac aaaggacatt aagtaaagac agataaacgt
     2221 agactaaaac gtggtcgcat cagggtgctg gcttttcaag ttccttaaga atggcctcaa
     2281 ttttctctat acactcagtt ggaacacgag acctgtccag gttaagcacc attttatcgc
     2341 ccttatacaa tactgtcgct ccaggagcaa actgatgtcg tgagcttaaa ctagttcttg
     2401 atgcagatga cgttttaagc acagaagtta aaagagtgat aacttcttca gcttcaaata
     2461 tcaccccagc ttttttctgc tcatgaaggt tagatgcctg ctgcttaagt aattcctctt
     2521 tatctgtaaa ggctttttga agtgcatcac ctgaccgggc agatagttca ccggggtgag
     2581 aaaaaagagc aacaactgat ttaggcaatt tggcggtgtt gatacagcgg gtaataatct
     2641 tacgtgaaat attttccgca tcagccagcg cagaaatatt tccagcaaat tcattctgca
     2701 atcggcttgc ataacgctga ccacgttcat aagcacttgt tgggcgataa tcgttaccca
     2761 atctggataa tgcagccatc tgctcatcat ccagctcgcc aaccagaaca cgataatcac
     2821 tttcggtaag tgcagcagct ttacgacggc gactcccatc ggcaatttct atgacaccag
     2881 atactcttcg accgaacgcc ggtgtctgtt gaccagtcag tagaaaagaa gggatgagat
     2941 catccagtgc gtcctcagta agcagctcct ggtcacgttc attacctgac catacccgag
     3001 aggtcttctc aacactatca ccccggagca cttcaagagt aaacttcaca tcccgaccac
     3061 atacaggcaa agtaatggca ttaccgcgag ccattactcc tacgcgcgca attaacgaat
     3121 ccaccatcgg ggcagctggt gtcgataacg aagtatcttc aaccggttga gtattgagcg
     3181 tatgttttgg aataacaggc gcacgcttca ttatctaatc tcccagcgtg gtttaatcag
     3241 acgatcgaaa atttcattgc agacaggttc ccaaatagaa agagcatttc tccaggcacc
     3301 agttgaagag cgttgatcaa tggcctgttc aaaaacagtt ctcatccgga tctgaccttt
     3361 accaacttca tccgtttcac gtacaacatt ttttagaacc atgcttcccc aggcatcccg
     3421 aatttgctcc tccatccacg gggactgaga gccattacta ttgctgtatt tggtaagcaa
     3481 aatacgtaca tcaggctcga accctttaag atcaacgttc ttgagcagat cacgaagcat
     3541 atcgaaaaac tgcagtgcgg aggtgtagtc aaacaactca gcaggcgtgg gaacaatcag
     3601 cacatcagca gcacatacga cattaatcgt gccgataccc aggttaggcg cgctgtcaat
     3661 aactatgaca tcatagtcat gagcaacagt ttcaatggcc agtcggagca tcaggtgtgg
     3721 atcggtgggc agtttacctt catcaaattt gcccattaac tcagtttcaa tacggtgcag
     3781 agccagacag gaaggaataa tgtcaagccc cggccagcaa gtgggcttta ttgcataagt
     3841 gacatcgtcc ttttccccaa gatagaaagg caggagagtg tcttctgcat gaatatgaag
     3901 atctggtacc catccgtgat acattgaggc tgttccctgg gggtcgttac cttccacgag
     3961 caaaacacgt agccccttca gagccagatc ctgagcaaga tgaacagaaa ctgaggtttt
     4021 gtaaacgcct cctttatggg cagcaacccc gatcaccggt ggaaatacgt cttcagcacg
     4081 tcgcaatcgc gtaccaaaca catcacgcat atgattaatt tgttcaattg tataaccaac
     4141 acgttgctca acccgtcctc gaatttccat atccgggtgc ggtagtcgcc ctgctttctc
     4201 ggcatctctg atagcctgag aagaaacccc aactaaatcc gctgcttcac ctattctcca
     4261 gcgccgggtt attttcctcg cttccgggct gtcatcatta aactgtgcaa tggcgatagc
     4321 cttcgtcatt tcatgaccag cgtttatgca ctggttaagt gtttccatga gtttcattct
     4381 gaacatcctt taatcattgc tttgcgtttt tttattaaat cttgcaattt actgcaaagc
     4441 aacaacaaaa tcgcaaagtc atcaaaaaac cgcaaagttg tttaaaataa gagcaacact
     4501 acaaaaggag ataagaagag cacatacctc agtcacttat tatcactagc gctcgccgca
     4561 gccgtgtaac cgagcatagc gagcgaactg gcgaggaagc aaagaagaac tgttctgtca
     4621 gatagctctt acgctcagcg caagaagaaa tatccaccgt gggaaaaact ccaggtagag
     4681 gtacacacgc ggatagccaa ttcagagtaa taaactgtga taatcaaccc tcatcaatga
     4741 tgacgaacta acccccgata tcaggtcaca tgacgaaggg aaagagaagg aaatcaactg
     4801 tgacaaactg ccctcaaatt tggcttcctt aaaaattaca gttcaaaaag tatgagaaaa
     4861 tccatgcagg ctgaaggaaa cagcaaaact gtgacaaatt accctcagta ggtcagaaca
     4921 aatgtgacga accaccctca aatctgtgac agataaccct cagactatcc tgtcgtcatg
     4981 gaagtgatat cgcggaagga aaatacgata tgagtcgtct ggcggccttt ctttttctca
     5041 atgtatgaga ggcgcattgg agttctgctg ttgatctcat taacacagac ctgcaggaag
     5101 cggcggcgga agtcaggcat acgctggtaa ctttgaggca gctggtaacg ctctatgatc
     5161 cagtcgattt tcagagagac gatgcctgag ccatccggct tacgatactg acacagggat
     5221 tcgtataaac gcatggcata cggattggtg atttcttttg tttcactaag ccgaaactgc
     5281 gtaaaccggt tctgtaaccc gataaagaag ggaatgagat atgggttgat atgtacactg
     5341 taaagccctc tggatggact gtgcgcacgt ttgataaacc aaggaaaaga ttcatagcct
     5401 ttttcatcgc cggcatcctc ttcagggcga taaaaaacca cttccttccc cgcgaaactc
     5461 ttcaatgcct gccgtatatc cttactggct tccgcagagg tcaatccgaa tatttcagca
     5521 tatttagcaa catggatctc gcagataccg tcatgttcct gtagggtgcc atcagatttt
     5581 ctgatctggt caacgaacag atacagcata cgtttttgat cccgggagag actatatgcc
     5641 gcctcagtga ggtcgtttga ctggacgatt cgcgggctat ttttacgttt cttgtgattg
     5701 ataaccgctg tttccgccat gacagatcca tgtgaagtgt gacaagtttt tagattgtca
     5761 cactaaataa aaaagagtca ataagcaggg ataactttgt gaaaaaacag cttcttctga
     5821 gggcaatttg tcacagggtt aagggcaatt tgtcacagac aggactgtca tttgagggtg
     5881 atttgtcaca ctgaaagggc aatttgtcac aacaccttct ctagaaccag catggataaa
     5941 ggcctacaag gcgctctaaa aaagaagatc taaaaactat aaaaaaaata attataaaaa
     6001 tatccccgtg gataagtgga taaccccaag ggaagttttt tcaggcatcg tgtgtaagca
     6061 gaatatataa gtgctgttcc ctggtgcttc ctcgctcact cgaccgggag ggttcgagaa
     6121 gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa
     6181 acaaggttta taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa
     6241 aacgggcgga aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc
     6301 aataggtgcg cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct
     6361 catctgtcag tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt
     6421 gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct
     6481 ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg
     6541 ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt
     6601 tccgcgaggt atccacaacg ccggcggccg gccgcggtgt ctcgcacacg gcttcgacgg
     6661 cgtttctggc gcgtttgcag ggccatagac ggccgccagc ccagcggcga gggcaaccag
     6721 ccgagggctt cgccctgtcg ctcgactgcg gcgagcacta ctggctgtaa aaggacagac
     6781 cacatcatgg ttctgtgttc attaggttgt tctgtccatt gctgacataa tccgctccac
     6841 ttcaacgtaa caccgcacga agatttctat tgttcctgaa ggcatattca aatcgttttc
     6901 gttaccgctt gcaggcatca tgacagaaca ctacttccta taaacgctac acaggctcct
     6961 gagattaata atgcggatct ctacgataat gggagatttt cccgactgtt tcgttcgctt
     7021 ctcagtggat aacagccagc ttctctgttt aacagacaaa aacagcatat ccactcagtt
     7081 ccacatttcc atataaaggc caaggcattt attctcagga taattgtttc agcatcgcaa
     7141 ccgcatcaga ctccggcatc gcaaactgca cccggtgccg ggcagccaca tccagcgcaa
     7201 aaaccttcgt gtagacttcc gttgaactga tggacttatg tcccatcagg ctttgcagaa
     7261 ctttcagcgg tataccggca tacagcatgt gcatcgcata ggaatggcgg aacgtatgtg
     7321 gtgtgaccgg aacagagaac gtcacaccgt cagcagcagc ggcggcaacc gcctccccaa
     7381 tccaggtcct gaccgttctg tccgtcactt cccagatccg cgctttctct gtccttcctg
     7441 tgcgacggtt acgccgctcc atgagcttat cgggtccact cgcgaacgca gaaaggccca
     7501 cccgaaggtg agccagtgtg attacatttg cggcctaact gtggccagtc cagttacgct
     7561 ggagtcacta gtatttaggt gacactatag aagcggccgc gacaacttgt ccacgtggaa
     7621 ttctaagctt aggatcccaa tgtgatgg
//
