LOCUS       Exported                7221 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBCPB+
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7221)
  AUTHORS   Groth AC, Olivares EC, Thyagarajan B, Calos MP
  TITLE     A phage integrase directs efficient site-specific integration in 
            human cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 May 23. 97(11):5995-6000.
  PUBMED    10801973
REFERENCE   2  (bases 1 to 7221)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7221
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    536..605
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     4144..4166
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    4391..4490
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(4594..4614)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4594..4612)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4633..4649)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4633..4649)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4646..4668)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4657..4673
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(4681..4711)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4726..4747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4864..4881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5035..5623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5115..5134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5843..5945
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5946..6605
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6012..6031)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7093..7198)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgcg ggccccccct cgaggtcgac gatgtaggtc acggtctcga
      541 agccgcggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac tccacctcac
      601 ccatctggtc catcatgatg aacgggtcga ggtggcggta gttgatcccg gcgaacgcgc
      661 ggcgcaccgg gaagccctcg ccctcgaaac cgctgggcgc ggtggtcacg gtgagcacgg
      721 gacgtgcgac ggcgtcggcg ggtgcggata cgcggggcag cgtcagcggg ttctcgacgg
      781 tcacggcggg catgtcgacg gtatcgataa gcttgggccc ctcgagggat cctctagagc
      841 ggccgcctag agtcgaggcc gagtttgtca gaaagcagac caaacagcgg ttggaataat
      901 agcgagaaca gagaaatagc ggcaaaaata atacccgtat cacttttgct gatatggttg
      961 atgtcatgta gccaaatcgg gaaaaacggg aagtaggctc ccatgataaa aaagtaaaag
     1021 aaaaagaata aaccgaacat ccaaaagttt gtgtttttta aatagtacat aatggatttc
     1081 cttacgcgaa atacgggcag acatggcctg cccggttatt attatttttg acaccagacc
     1141 aactggtaat ggtagcgacc ggcgctcagc tgtaattccg ccgatactga cgggctccag
     1201 gagtcgtcgc caccaatccc catatggaaa ccgtcgatat tcagccatgt gccttcttcc
     1261 gcgtgcagca gatggcgatg gctggtttcc atcgttgctg ttgactgtag cggctgatgt
     1321 tgaactggaa gtcgccgcgc cactggtgtg ggccataatt caattcgcgc gtcccgcagc
     1381 gcagaccgtt ttcgctcggg aagacgtacg gggtatacat gtctgacaat ggcagatccc
     1441 agcggtcaaa acaggcggca gtaaggcggt cgggatagtt ttcttgcggc cctaatccga
     1501 gccagtttac ccgctctgct acctgcgcca gctggcagtt caggccaatc cgcgccggat
     1561 gcggtgtatc gctcgccact tcaacatcaa cggtaatcgc catttgacca ctaccatcaa
     1621 tccggtaggt tttccggctg ataaataagg ttttcccctg atgctgccac gcgtgagcgg
     1681 tcgtaatcag caccgcatca gcaagtgtat ctgccgtgca ctgcaacaac gctgcttcgg
     1741 cctggtaatg gcccgccgcc ttccagcgtt cgacccaggc gttagggtca atgcgggtcg
     1801 cttcacttac gccaatgtcg ttatccagcg gtgcacgggt gaactgatcg cgcagcggcg
     1861 tcagcagttg ttttttatcg ccaatccaca tctgtgaaag aaagcctgac tggcggttaa
     1921 attgccaacg cttattaccc agctcgatgc aaaaatccat ttcgctggtg gtcagatgcg
     1981 ggatggcgtg ggacgcggcg gggagcgtca cactgaggtt ttccgccaga cgccactgct
     2041 gccaggcgct gatgtgcccg gcttctgacc atgcggtcgc gttcggttgc actacgcgta
     2101 ctgtgagcca gagttgcccg gcgctctccg gctgcggtag ttcaggcagt tcaatcaact
     2161 gtttaccttg tggagcgaca tccagaggca cttcaccgct tgccagcggc ttaccatcca
     2221 gcgccaccat ccagtgcagg agctcgttat cgctatgacg gaacaggtat tcgctggtca
     2281 cttcgatggt ttgcccggat aaacggaact ggaaaaactg ctgctggtgt tttgcttccg
     2341 tcagcgctgg atgcggcgtg cggtcggcaa agaccagacc gttcatacag aactggcgat
     2401 cgttcggcgt atcgccaaaa tcaccgccgt aagccgacca cgggttgccg ttttcatcat
     2461 atttaatcag cgactgatcc acccagtccc agacgaagcc gccctgtaaa cggggatact
     2521 gacgaaacgc ctgccagtat ttagcgaaac cgccaagact gttacccatc gcgtgggcgt
     2581 attcgcaaag gatcagcggg cgcgtctctc caggtagcga aagccatttt ttgatggacc
     2641 atttcggcac agccgggaag ggctggtctt catccacgcg cgcgtacatc gggcaaataa
     2701 tatcggtggc cgtggtgtcg gctccgccgc cttcatactg caccgggcgg gaaggatcga
     2761 cagatttgat ccagcgatac agcgcgtcgt gattagcgcc gtggcctgat tcattcccca
     2821 gcgaccagat gatcacactc gggtgattac gatcgcgctg caccattcgc gttacgcgtt
     2881 cgctcatcgc cggtagccag cgcggatcat cggtcagacg attcattggc accatgccgt
     2941 gggtttcaat attggcttca tccaccacat acaggccgta gcggtcgcac agcgtgtacc
     3001 acagcggatg gttcggataa tgcgaacagc gcacggcgtt aaagttgttc tgcttcatca
     3061 gcaggatatc ctgcaccatc gtctgctcat ccatgacctg accatgcaga ggatgatgct
     3121 cgtgacggtt aacgcctcga atcagcaacg gcttgccgtt cagcagcagc agaccatttt
     3181 caatccgcac ctcgcggaaa ccgacatcgc aggcttctgc ttcaatcagc gtgccgtcgg
     3241 cggtgtgcag ttcaaccacc gcacgataga gattcgggat ttcggcgctc cacagtttcg
     3301 ggttttcgac gttcagacgt agtgtgacgc gatcggcata accaccacgc tcatcgataa
     3361 tttcaccgcc gaaaggcgcg gtgccgctgg cgacctgcgt ttcaccctgc cataaagaaa
     3421 ctgttacccg taggtagtca cgcaactcgc cgcacatctg aacttcagcc tccagtacag
     3481 cgcggctgaa atcatcatta aagcgagtgg caacatggaa atcgctgatt tgtgtagtcg
     3541 gtttatgcag caacgagacg tcacggaaaa tgccgctcat ccgccacata tcctgatctt
     3601 ccagataact gccgtcactc caacgcagca ccatcaccgc gaggcggttt tctccggcgc
     3661 gtaaaaatgc gctcaggtca aattcagacg gcaaacgact gtcctggccg taaccgaccc
     3721 agcgcccgtt gcaccacaga tgaaacgccg agttaacgcc atcaaaaata attcgcgtct
     3781 ggccttcctg tagccagctt tcatcaacat taaatgtgag cgagtaacaa cccgtcggat
     3841 tctccgtggg aacaaacggc ggattgaccg taatgggata ggttacgttg gtgtagatgg
     3901 gcgcatcgta accgtgcatc tgccagtttg aggggacgac gacagtatcg gcctcaggaa
     3961 gatcgcactc cagccagctt tccggcaccg cttctggtgc cggaaaccag gcaaagcgcc
     4021 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4081 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     4141 tttcccagtc acgacgttgt aaaacgacgg gatcgcgctt gagcagctcc ttgctggtgt
     4201 ccagaccaat gcctcccaga ccggcaacga aaatcacgtt cttgttggtc aaagtaaacg
     4261 acatggtgac ttcttttttg ctttagcagg ctctttcgat ccccgggaat tgcggccgcc
     4321 gactagtact gacggacaca ccgaagcccc ggcggcaacc ctcagcggat gccccggggc
     4381 ttcacgtttt cccaggtcag aagcggtttt cgggagtagt gccccaactg gggtaacctt
     4441 tgagttctct cagttggggg cgtagggtcg ccgacatgac acaaggggtt gtgaccgggg
     4501 tggacacgta cgcgggtgct tacgaccgtc agtcgcgcga gcgcgactag ttctagagcg
     4561 gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     4621 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4681 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4741 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4801 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4861 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4921 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4981 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5041 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5101 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5161 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5221 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5281 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5341 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5401 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5461 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5521 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5581 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5641 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5701 attatcaaaa aggatcttca cctagatcct tttcgaccga ataaatacct gtgacggaag
     5761 atcacttcgc agaataaata aatcctggtg tccctgttga taccgggaag ccctgggcca
     5821 acttttggcg aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg
     5881 aaataagatc actaccgggc gtattttttg agttgtcgag attttcagga gctaaggaag
     5941 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     6001 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     6061 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     6121 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attacgtatg gcaatgaaag
     6181 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     6241 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     6301 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     6361 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     6421 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     6481 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     6541 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     6601 cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggttgctac gcctgaataa
     6661 gtgataataa gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc
     6721 agggcagggt cgttaaatag ccgcttatgt ctattgctgg tttaccggtt tattgactac
     6781 cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc aggctctccc
     6841 cgtggaggta ataattgacg atatgatcct ttttttctga tcaaaagtgc tcatcattgg
     6901 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6961 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7021 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7081 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcaag ggttattgtc
     7141 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7201 catttccccg aaaagtgcca c
//
