LOCUS       40924_9251        8181 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCC2FOS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8181)
  AUTHORS   EPICENTRE Biotechnologies.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2007) 726 Post Road, Madison, WI 53713, USA
REFERENCE   2  (bases 1 to 8181)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8181)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (23-AUG-2007) 726 Post Road, Madison, WI 53713, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     288..304
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        311..329
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(485..501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(509..525)
                     /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(533..563)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(578..599)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(822..1478)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1479..1581)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      2439..3053
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      3129..3348
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             3439..4191
                     /codon_start=1
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4197..4447
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             4773..5945
                     /codon_start=1
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             5948..6916
                     /codon_start=1
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    6992..7465
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    complement(7725..8123)
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(8141..8174)
                     /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 cagctgcgca actgttggga
      181 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      241 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      301 cagtgaattg taatacgact cactataggg cgaattcgag ctcggtaccc ggggatccca
      361 cgtacaacga cacctagacc acgtgttcct aggctgtttc ctggtgggat cctctagagt
      421 cgacctgcag gcatgcaagc ttgagtattc tatagtctca cctaaatagc ttggcgtaat
      481 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
      541 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
      601 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
      661 gaatcggcca acgcgaaccc cttgcggccg cccgggccgt cgaccaattc tcatgtttga
      721 cagcttatca tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcaa
      781 ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca
      841 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
      901 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
      961 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     1021 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     1081 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     1141 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     1201 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
     1261 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     1321 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     1381 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     1441 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     1501 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     1561 gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg
     1621 gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt
     1681 tattcgcgat aagctcatgg agcggcgtaa ccgtcgcaca ggaaggacag agaaagcgcg
     1741 gatctgggaa gtgacggaca gaacggtcag gacctggatt ggggaggcgg ttgccgccgc
     1801 tgctgctgac ggtgtgacgt tctctgttcc ggtcacacca catacgttcc gccattccta
     1861 tgcgatgcac atgctgtatg ccggtatacc gctgaaagtt ctgcaaagcc tgatgggaca
     1921 taagtccatc agttcaacgg aagtctacac gaaggttttt gcgctggatg tggctgcccg
     1981 gcaccgggtg cagtttgcga tgccggagtc tgatgcggtt gcgatgctga aacaattatc
     2041 ctgagaataa atgccttggc ctttatatgg aaatgtggaa ctgagtggat atgctgtttt
     2101 tgtctgttaa acagagaagc tggctgttat ccactgagaa gcgaacgaaa cagtcgggaa
     2161 aatctcccat tatcgtagag atccgcatta ttaatctcag gagcctgtgt agcgtttata
     2221 ggaagtagtg ttctgtcatg atgcctgcaa gcggtaacga aaacgatttg aatatgcctt
     2281 caggaacaat agaaatcttc gtgcggtgtt acgttgaagt ggagcggatt atgtcagcaa
     2341 tggacagaac aacctaatga acacagaacc atgatgtggt ctgtcctttt acagccagta
     2401 gtgctcgccg cagtcgagcg acagggcgaa gccctcggct ggttgccctc gccgctgggc
     2461 tggcggccgt ctatggccct gcaaacgcgc cagaaacgcc gtcgaagccg tgtgcgagac
     2521 accgcggccg gccgccggcg ttgtggatac ctcgcggaaa acttggccct cactgacaga
     2581 tgaggggcgg acgttgacac ttgaggggcc gactcacccg gcgcggcgtt gacagatgag
     2641 gggcaggctc gatttcggcc ggcgacgtgg agctggccag cctcgcaaat cggcgaaaac
     2701 gcctgatttt acgcgagttt cccacagatg atgtggacaa gcctggggat aagtgccctg
     2761 cggtattgac acttgagggg cgcgactact gacagatgag gggcgcgatc cttgacactt
     2821 gaggggcaga gtgctgacag atgaggggcg cacctattga catttgaggg gctgtccaca
     2881 ggcagaaaat ccagcatttg caagggtttc cgcccgtttt tcggccaccg ctaacctgtc
     2941 ttttaacctg cttttaaacc aatatttata aaccttgttt ttaaccaggg ctgcgccctg
     3001 tgcgcgtgac cgcgcacgcc gaaggggggt gccccccctt ctcgaaccct cccggtcgag
     3061 tgagcgagga agcaccaggg aacagcactt atatattctg cttacacacg atgcctgaaa
     3121 aaacttccct tggggttatc cacttatcca cggggatatt tttataatta ttttttttat
     3181 agtttttaga tcttcttttt tagagcgcct tgtaggcctt tatccatgct ggttctagag
     3241 aaggtgttgt gacaaattgc cctttcagtg tgacaaatca ccctcaaatg acagtcctgt
     3301 ctgtgacaaa ttgcccttaa ccctgtgaca aattgccctc agaagaagct gttttttcac
     3361 aaagttatcc ctgcttattg actctttttt atttagtgtg acaatctaaa aacttgtcac
     3421 acttcacatg gatctgtcat ggcggaaaca gcggttatca atcacaagaa acgtaaaaat
     3481 agcccgcgaa tcgtccagtc aaacgacctc actgaggcgg catatagtct ctcccgggat
     3541 caaaaacgta tgctgtatct gttcgttgac cagatcagaa aatctgatgg caccctacag
     3601 gaacatgacg gtatctgcga gatccatgtt gctaaatatg ctgaaatatt cggattgacc
     3661 tctgcggaag ccagtaagga tatacggcag gcattgaaga gtttcgcggg gaaggaagtg
     3721 gttttttatc gccctgaaga ggatgccggc gatgaaaaag gctatgaatc ttttccttgg
     3781 tttatcaaac gtgcgcacag tccatccaga gggctttaca gtgtacatat caacccatat
     3841 ctcattccct tctttatcgg gttacagaac cggtttacgc agtttcggct tagtgaaaca
     3901 aaagaaatca ccaatccgta tgccatgcgt ttatacgaat ccctgtgtca gtatcgtaag
     3961 ccggatggct caggcatcgt ctctctgaaa atcgactgga tcatagagcg ttaccagctg
     4021 cctcaaagtt accagcgtat gcctgacttc cgccgccgct tcctgcaggt ctgtgttaat
     4081 gagatcaaca gcagaactcc aatgcgcctc tcatacattg agaaaaagaa aggccgccag
     4141 acgactcata tcgtattttc cttccgcgat atcacttcca tgacgacagg atagtctgag
     4201 ggttatctgt cacagatttg agggtggttc gtcacatttg ttctgaccta ctgagggtaa
     4261 tttgtcacag ttttgctgtt tccttcagcc tgcatggatt ttctcatact ttttgaactg
     4321 taatttttaa ggaagccaaa tttgagggca gtttgtcaca gttgatttcc ttctctttcc
     4381 cttcgtcatg tgacctgata tcgggggtta gttcgtcatc attgatgagg gttgattatc
     4441 acagtttatt actctgaatt ggctatccgc gtgtgtacct ctacctggag tttttcccac
     4501 ggtggatatt tcttcttgcg ctgagcgtaa gagctatctg acagaacagt tcttctttgc
     4561 ttcctcgcca gttcgctcgc tatgctcggt tacacggctg cggcgagcgc tagtgataat
     4621 aagtgactga ggtatgtgct cttcttatct ccttttgtag tgttgctctt attttaaaca
     4681 actttgcggt tttttgatga ctttgcgatt ttgttgttgc tttgcagtaa attgcaagat
     4741 ttaataaaaa aacgcaaagc aatgattaaa ggatgttcag aatgaaactc atggaaacac
     4801 ttaaccagtg cataaacgct ggtcatgaaa tgacgaaggc tatcgccatt gcacagttta
     4861 atgatgacag cccggaagcg aggaaaataa cccggcgctg gagaataggt gaagcagcgg
     4921 atttagttgg ggtttcttct caggctatca gagatgccga gaaagcaggg cgactaccgc
     4981 acccggatat ggaaattcga ggacgggttg agcaacgtgt tggttataca attgaacaaa
     5041 ttaatcatat gcgtgatgtg tttggtacgc gattgcgacg tgctgaagac gtatttccac
     5101 cggtgatcgg ggttgctgcc cataaaggtg gcgtttacaa aacctcagtt tctgttcatc
     5161 ttgctcagga tctggctctg aaggggctac gtgttttgct cgtggaaggt aacgaccccc
     5221 agggaacagc ctcaatgtat cacggatggg taccagatct tcatattcat gcagaagaca
     5281 ctctcctgcc tttctatctt ggggaaaagg acgatgtcac ttatgcaata aagcccactt
     5341 gctggccggg gcttgacatt attccttcct gtctggctct gcaccgtatt gaaactgagt
     5401 taatgggcaa atttgatgaa ggtaaactgc ccaccgatcc acacctgatg ctccgactgg
     5461 ccattgaaac tgttgctcat gactatgatg tcatagttat tgacagcgcg cctaacctgg
     5521 gtatcggcac gattaatgtc gtatgtgctg ctgatgtgct gattgttccc acgcctgctg
     5581 agttgtttga ctacacctcc gcactgcagt ttttcgatat gcttcgtgat ctgctcaaga
     5641 acgttgatct taaagggttc gagcctgatg tacgtatttt gcttaccaaa tacagcaata
     5701 gtaatggctc tcagtccccg tggatggagg agcaaattcg ggatgcctgg ggaagcatgg
     5761 ttctaaaaaa tgttgtacgt gaaacggatg aagttggtaa aggtcagatc cggatgagaa
     5821 ctgtttttga acaggccatt gatcaacgct cttcaactgg tgcctggaga aatgctcttt
     5881 ctatttggga acctgtctgc aatgaaattt tcgatcgtct gattaaacca cgctgggaga
     5941 ttagataatg aagcgtgcgc ctgttattcc aaaacatacg ctcaatactc aaccggttga
     6001 agatacttcg ttatcgacac cagctgcccc gatggtggat tcgttaattg cgcgcgtagg
     6061 agtaatggct cgcggtaatg ccattacttt gcctgtatgt ggtcgggatg tgaagtttac
     6121 tcttgaagtg ctccggggtg atagtgttga gaagacctct cgggtatggt caggtaatga
     6181 acgtgaccag gagctgctta ctgaggacgc actggatgat ctcatccctt cttttctact
     6241 gactggtcaa cagacaccgg cgttcggtcg aagagtatct ggtgtcatag aaattgccga
     6301 tgggagtcgc cgtcgtaaag ctgctgcact taccgaaagt gattatcgtg ttctggttgg
     6361 cgagctggat gatgagcaga tggctgcatt atccagattg ggtaacgatt atcgcccaac
     6421 aagtgcttat gaacgtggtc agcgttatgc aagccgattg cagaatgaat ttgctggaaa
     6481 tatttctgcg ctggctgatg cggaaaatat ttcacgtaag attattaccc gctgtatcaa
     6541 caccgccaaa ttgcctaaat cagttgttgc tcttttttct caccccggtg aactatctgc
     6601 ccggtcaggt gatgcacttc aaaaagcctt tacagataaa gaggaattac ttaagcagca
     6661 ggcatctaac cttcatgagc agaaaaaagc tggggtgata tttgaagctg aagaagttat
     6721 cactctttta acttctgtgc ttaaaacgtc atctgcatca agaactagtt taagctcacg
     6781 acatcagttt gctcctggag cgacagtatt gtataagggc gataaaatgg tgcttaacct
     6841 ggacaggtct cgtgttccaa ctgagtgtat agagaaaatt gaggccattc ttaaggaact
     6901 tgaaaagcca gcaccctgat gcgaccacgt tttagtctac gtttatctgt ctttacttaa
     6961 tgtcctttgt tacaggccag aaagcataac tggcctgaat attctctctg ggcccactgt
     7021 tccacttgta tcgtcggtct gataatcaga ctgggaccac ggtcccactc gtatcgtcgg
     7081 tctgattatt agtctgggac cacggtccca ctcgtatcgt cggtctgatt attagtctgg
     7141 gaccacggtc ccactcgtat cgtcggtctg ataatcagac tgggaccacg gtcccactcg
     7201 tatcgtcggt ctgattatta gtctgggacc atggtcccac tcgtatcgtc ggtctgatta
     7261 ttagtctggg accacggtcc cactcgtatc gtcggtctga ttattagtct ggaaccacgg
     7321 tcccactcgt atcgtcggtc tgattattag tctgggacca cggtcccact cgtatcgtcg
     7381 gtctgattat tagtctggga ccacgatccc actcgtgttg tcggtctgat tatcggtctg
     7441 ggaccacggt cccacttgta ttgtcgatca gactatcagc gtgagactac gattccatca
     7501 atgcctgtca agggcaagta ttgacatgtc gtcgtaacct gtagaacgga gtaacctcgg
     7561 tgtgcggttg tatgcctgct gtggattgct gctgtgtcct gcttatccac aacattttgc
     7621 gcacggttat gtggacaaaa tacctggtta cccaggccgt gccggcacgt taaccgggct
     7681 gcatccgatg caagtgtgtc gctgtcgacg agctcgcgag ctcggacatg aggttgcccc
     7741 gtattcagtg tcgctgattt gtattgtctg aagttgtttt tacgttaagt tgatgcagat
     7801 caattaatac gatacctgcg tcataattga ttatttgacg tggtttgatg gcctccacgc
     7861 acgttgtgat atgtagatga taatcattat cactttacgg gtcctttccg gtgatccgac
     7921 aggttacggg gcggcgacct cgcgggtttt cgctatttat gaaaattttc cggtttaagg
     7981 cgtttccgtt cttcttcgtc ataacttaat gtttttattt aaaataccct ctgaaaagaa
     8041 aggaaacgac aggtgctgaa agcgagcttt ttggcctctg tcgtttcctt tctctgtttt
     8101 tgtccgtgga atgaacaatg gaagtccgag ctcatcgcta ataacttcgt atagcataca
     8161 ttatacgaag ttatattcga t
//
