LOCUS       40924_40717        6900 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Large-insert cloning vector pSMART VC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  AUTHORS   Wild J, Hradecna Z, Szybalski W.
  TITLE     Conditionally amplifiable BACs: switching from single-copy to 
            high-copy vectors and genomic clones
  JOURNAL   Genome Res. 12 (9), 1434-1444 (2002)
  PUBMED    12213781
REFERENCE   2  (bases 1 to 6900)
  AUTHORS   Godiska R, Mead DA.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAY-2004) Lucigen Corp., 2120 W. Greenview Dr. #9, 
            Middleton, WI 53562, USA
REFERENCE   3  (bases 1 to 6900)
  AUTHORS   Godiska R, Mead DA.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2005) Lucigen Corp., 2120 W. Greenview Dr. #9, 
            Middleton, WI 53562, USA
REFERENCE   4  (bases 1 to 6900)
  AUTHORS   Godiska R, Mead DA.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JUN-2005) Lucigen Corp., 2120 W. Greenview Dr. #9, 
            Middleton, WI 53562, USA
REFERENCE   5  (bases 1 to 6900)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 6900)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome 
            Res."; date: "2002"; volume: "12"; issue: "9"; pages: "1434-1444"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-MAY-2004) Lucigen Corp., 2120 W. Greenview Dr. #9, Middleton, WI
            53562, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (13-JAN-2005) Lucigen Corp., 2120 W. Greenview Dr. #9, Middleton, WI
            53562, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: "Submitted 
            (27-JUN-2005) Lucigen Corp., 2120 W. Greenview Dr. #9, Middleton, WI
            53562, USA"
COMMENT     SGRef: number: 5; type: "Journal Article"
COMMENT     On Jun 27, 2005 this sequence version replaced AY643800.2.
FEATURES             Location/Qualifiers
     source          1..6900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(28..57)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     protein_bind    79..112
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    176..231
                     /label=multiple cloning site
                     /note="multiple cloning site"
     protein_bind    complement(273..306)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      325..356
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     misc_feature    405..803
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     promoter        825..927
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             928..1584
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     misc_feature    complement(1858..2331)
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(2407..3375)
                     /codon_start=1
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(3378..4550)
                     /codon_start=1
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    complement(4876..5126)
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(5132..5884)
                     /codon_start=1
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      complement(5975..6194)
                     /direction=LEFT
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      complement(6268..6897)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 aaggaatatt cagcaatttg cccgtgccga agaaaggccc acccgtgaag gtgagccagt
       61 gagttgattg ctacgtaaat aacttcgtat agcatacatt atacgaagtt atggactagg
      121 cgcgccagaa gagagaaaga aggaaagcgg ccgccagggt tttcccagtc acgacaagct
      181 tgcatgcgat acgtatcgtt aacgatggat ccgacgcacg tgcgaattcc tctagaggtt
      241 cctagttgta actgccaggg cggccacttg acataacttc gtatagcata cattatacga
      301 agttatgttt aaacattagc agaaagtcaa aagcctccga ccggaggctt ttgactaaaa
      361 cttcccttgg ggttatcatt gggccgagac cgcctgaaga ggacttccat tgttcattcc
      421 acggacaaaa acagagaaag gaaacgacag aggccaaaaa gctcgctttc agcacctgtc
      481 gtttcctttc ttttcagagg gtattttaaa taaaaacatt aagttatgac gaagaagaac
      541 ggaaacgcct taaaccggaa aattttcata aatagcgaaa acccgcgagg tcgccgcccc
      601 gtaacctgtc ggatcaccgg aaaggacccg taaagtgata atgattatca tctacatatc
      661 acaacgtgcg tggaggccat caaaccacgt caaataatca attatgacgc aggtatcgta
      721 ttaattgatc tgcatcaact taacgtaaaa gcaacttcag acaatacaaa tcagcgacac
      781 tgaatacggg gcaacctcat gtcgcctgaa gagtgagacc gccctgatcg gcacgtaaga
      841 ggttccgact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
      901 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
      961 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     1021 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     1081 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1141 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1201 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1261 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1321 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1381 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1441 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1501 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1561 tgcgatgagt ggcagggcgg ggcgtaaaaa tgtaatcacc tggctcacct tcgggtgggc
     1621 ctttcacact tgcatcggat gcagcccggt gaacgtgccg gcacggcctg ggtaaccagg
     1681 tattttgtcc acataaccgt gcgcaaaatg ttgtggataa gcaggacaca gcagcaatcc
     1741 acagcaggca tacaaccgca caccgaggtt actccgttct acaggttacg acgacatgtc
     1801 aatacttgcc cttgacaggc attgatggaa tcgtagtctc acgctgatag tctgatcgac
     1861 aatacaagtg ggaccgtggt cccagaccga taatcagacc gacaacacga gtgggatcgt
     1921 ggtcccagac taataatcag accgacgata cgagtgggac cgtggtccca gactaataat
     1981 cagaccgacg atacgagtgg gaccgtggtt ccagactaat aatcagaccg acgatacgag
     2041 tgggaccgtg gtcccagact aataatcaga ccgacgatac gagtgggacc atggtcccag
     2101 actaataatc agaccgacga tacgagtggg accgtggtcc cagtctgatt atcagaccga
     2161 cgatacgagt gggaccgtgg tcccagacta ataatcagac cgacgatacg agtgggaccg
     2221 tggtcccaga ctaataatca gaccgacgat acgagtggga ccgtggtccc agtctgatta
     2281 tcagaccgac gatacaagtg gaacagtggg cccagagaga atattcaggc cagttatgct
     2341 ttctggcctg taacaaagga cattaagtaa agacagataa acgtagacta aaacgtggtc
     2401 gcatcagggt gctggctttt caagttcctt aagaatggcc tcaattttct ctatacactc
     2461 agttggaaca cgagacctgt ccaggttaag caccatttta tcgcccttat acaatactgt
     2521 cgctccagga gcaaactgat gtcgtgagct taaactagtt cttgatgcag atgacgtttt
     2581 aagcacagaa gttaaaagag tgataacttc ttcagcttca aatatcaccc cagctttttt
     2641 ctgctcatga aggttagatg cctgctgctt aagtaattcc tctttatctg taaaggcttt
     2701 ttgaagtgca tcacctgacc gggcagatag ttcaccgggg tgagaaaaaa gagcaacaac
     2761 tgatttaggc aatttggcgg tgttgataca gcgggtaata atcttacgtg aaatattttc
     2821 cgcatcagcc agcgcagaaa tatttccagc aaattcattc tgcaatcggc ttgcataacg
     2881 ctgaccacgt tcataagcac ttgttgggcg ataatcgtta cccaatctgg ataatgcagc
     2941 catctgctca tcatccagct cgccaaccag aacacgataa tcactttcgg taagtgcagc
     3001 agctttacga cggcgactcc catcggcaat ttctatgaca ccagatactc ttcgaccgaa
     3061 cgccggtgtc tgttgaccag tcagtagaaa agaagggatg agatcatcca gtgcgtcctc
     3121 agtaagcagc tcctggtcac gttcattacc tgaccatacc cgagaggtct tctcaacact
     3181 atcaccccgg agcacttcaa gagtaaactt cacatcccga ccacatacag gcaaagtaat
     3241 ggcattaccg cgagccatta ctcctacgcg cgcaattaac gaatccacca tcggggcagc
     3301 tggtgtcgat aacgaagtat cttcaaccgg ttgagtattg agcgtatgtt ttggaataac
     3361 aggcgcacgc ttcattatct aatctcccag cgtggtttaa tcagacgatc gaaaatttca
     3421 ttgcagacag gttcccaaat agaaagagca tttctccagg caccagttga agagcgttga
     3481 tcaatggcct gttcaaaaac agttctcatc cggatctgac ctttaccaac ttcatccgtt
     3541 tcacgtacaa cattttttag aaccatgctt ccccaggcat cccgaatttg ctcctccatc
     3601 cacggggact gagagccatt actattgctg tatttggtaa gcaaaatacg tacatcaggc
     3661 tcgaaccctt taagatcaac gttcttgagc agatcacgaa gcatatcgaa aaactgcagt
     3721 gcggaggtgt agtcaaacaa ctcagcaggc gtgggaacaa tcagcacatc agcagcacat
     3781 acgacattaa tcgtgccgat acccaggtta ggcgcgctgt caataactat gacatcatag
     3841 tcatgagcaa cagtttcaat ggccagtcgg agcatcaggt gtggatcggt gggcagttta
     3901 ccttcatcaa atttgcccat taactcagtt tcaatacggt gcagagccag acaggaagga
     3961 ataatgtcaa gccccggcca gcaagtgggc tttattgcat aagtgacatc gtccttttcc
     4021 ccaagataga aaggcaggag agtgtcttct gcatgaatat gaagatctgg tacccatccg
     4081 tgatacattg aggctgttcc ctgggggtcg ttaccttcca cgagcaaaac acgtagcccc
     4141 ttcagagcca gatcctgagc aagatgaaca gaaactgagg ttttgtaaac gccaccttta
     4201 tgggcagcaa ccccgatcac cggtggaaat acgtcttcag cacgtcgcaa tcgcgtacca
     4261 aacacatcac gcatatgatt aatttgttca attgtataac caacacgttg ctcaacccgt
     4321 cctcgaattt ccatatccgg gtgcggtagt cgccctgctt tctcggcatc tctgatagcc
     4381 tgagaagaaa ccccaactaa atccgctgct tcacctattc tccagcgccg ggttattttc
     4441 ctcgcttccg ggctgtcatc attaaactgt gcaatggcga tagccttcgt catttcatga
     4501 ccagcgttta tgcactggtt aagtgtttcc atgagtttca ttctgaacat cctttaatca
     4561 ttgctttgcg tttttttatt aaatcttgca atttactgca aagcaacaac aaaatcgcaa
     4621 agtcatcaaa aaaccgcaaa gttgtttaaa ataagagcaa cactacaaaa ggagataaga
     4681 agagcacata cctcagtcac ttattatcac tagcgctcgc cgcagccgtg taaccgagca
     4741 tagcgagcga actggcgagg aagcaaagaa gaactgttct gtcagatagc tcttacgctc
     4801 agcgcaagaa gaaatatcca ccgtgggaaa aactccaggt agaggtacac acgcggatag
     4861 ccaattcaga gtaataaact gtgataatca accctcatca atgatgacga actaaccccc
     4921 gatatcaagt cacatgacga agggaaagag aaggaaatca actgtgacaa actgccctca
     4981 aatttggctt ccttaaaaat tacagttcaa aaagtatgag aaaatccatg caggctgaag
     5041 gaaacagcaa aactgtgaca aattaccctc agtaggtcag aacaaatgtg acgaaccacc
     5101 ctcaaatctg tgacagataa ccctcagact atcctgtcgt catggaagtg atatcgcgga
     5161 aggaaaatac gatatgagtc gtctggcggc ctttcttttt ctcaatgtat gagaggcgca
     5221 ttggagttct gctgttgatc tcattaacac agacctgcag gaagcggcgg cggaagtcag
     5281 gcatacgctg gtaactttga ggcagctggt aacgctctat gatccagtcg attttcagag
     5341 agacgatgcc tgagccatcc ggcttacgat actgacacag ggattcgtat aaacgcatgg
     5401 catacggatt ggtgatttct tttgtttcac taagccgaaa ctgcgtaaac cggttctgta
     5461 acccgataaa gaagggaatg agatatgggt tgatatgtac actgtaaagc cctctggatg
     5521 gactgtgcgc acgtttgata aaccaaggaa aagattcata gcctttttca tcgccggcat
     5581 cctcttcagg gcgataaaaa accacttcct tccccgcgaa actcttcaat gcctgccgta
     5641 tatccttact ggcttccgca gaggtcaatc cgaatatttc agcatattta gcaacatgga
     5701 tctcgcagat accgtcatgt tcctgtaggg tgccatcaga ttttctgatc tggtcaacga
     5761 acagatacag catacgtttt tgatcccggg agagactata tgccgcctca gtgaggtcgt
     5821 ttgactggac gattcgcggg ctatttttac gtttcttgtg attgataacc gctgtttccg
     5881 ccatgacaga tccatgtgaa gtgtgacaag tttttagatt gtcacactaa ataaaaaaga
     5941 gtcaataagc agggataact ttgtgaaaaa acagcttctt ctgagggcaa tttgtcacag
     6001 ggttaagggc aatttgtcac agacaggact gtcatttgag ggtgatttgt cacactgaaa
     6061 gggcaatttg tcacaacacc ttctctagaa ccagcatgga taaaggccta caaggcgctc
     6121 taaaaaagaa gatctaaaaa ctataaaaaa aataattata aaaatatccc cgtggataag
     6181 tggataaccc caagggaagt tttttcaggc atcgtgtgta agcagaatat ataagtgctg
     6241 ttccctggtg cttcctcgct cactcgaccg ggagggttcg agaagggggg gcacccccct
     6301 tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg tttataaata
     6361 ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg cggaaaccct
     6421 tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc
     6481 atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg
     6541 cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac atcatctgtg
     6601 ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag ctccacgtcg
     6661 ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag tcggcccctc
     6721 aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg aggtatccac
     6781 aacgccggcg gccggccgcg gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt
     6841 gcagggccat agacggccgc cagcccagcg gcgagggcaa ccagcccggt gagcgtcgga
//
