LOCUS       Exported                6230 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCMVInt
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6230)
  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 6230)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6230
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      2..457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     299..320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             745..1605
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(963..982)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    complement(1707..1740)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      1818..2406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2307..2326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2560..2577
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2907..2923
                     /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)."
     primer_bind     2912..2934
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     enhancer        2953..3256
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3257..3460
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3411..3431
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3457..3481
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3576..3594
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3576..3593
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3687..5504
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     primer_bind     complement(5567..5586)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5568..5586)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5973..6107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6010..6029)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6064..6083
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      421 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat
      481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      541 ccaggtcttg gactggtgag aacggcttgc tcggcagctt cgatgtgtgc tggagggaga
      601 ataaaggtct aagatgtgcg atagagggaa gtcgcattga attatgtgct gtgtagggat
      661 cgctggtatc aaatatgtgt gcccacccct ggcatgagac aataaccctg ataaatgctt
      721 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      781 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      841 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      901 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
      961 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1021 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1081 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1141 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1201 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1261 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     1321 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1381 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1441 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     1501 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1561 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1621 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1681 aagatccttt ttgataatct catgccataa cttcgtataa tgtatgctat acgaagttat
     1741 ggcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1801 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1861 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1921 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     1981 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2041 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2101 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2161 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     2221 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2281 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2341 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2401 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2461 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2521 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2581 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2641 agagcttgca attcgcgcgt ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2701 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2761 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     2821 aaaaataggc gtatacgagg ccctttcact cattaggcac cccaggcttt acactttata
     2881 gcttccggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
     2941 catcgtgcag gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     3001 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     3061 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     3121 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     3181 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     3241 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3301 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     3361 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     3421 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     3481 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     3541 cctccggact ctagcctagg cttttgcaaa aagctattta ggtgacacta tagaaggtac
     3601 gcctgcaggt accggtccgg aattcccggg tcgacgagct cactagtcgt agggtcgccg
     3661 acatgacaca aggggttgtg accggggtgg acacgtacgc gggtgcttac gaccgtcagt
     3721 cgcgcgagcg cgaaaattcg agcgcagcaa gcccagcgac acagcgtagc gccaacgaag
     3781 acaaggcggc cgaccttcag cgcgaagtcg agcgcgacgg gggccggttc aggttcgtcg
     3841 ggcatttcag cgaagcgccg ggcacgtcgg cgttcgggac ggcggagcgc ccggagttcg
     3901 aacgcatcct gaacgaatgc cgcgccgggc ggctcaacat gatcattgtc tatgacgtgt
     3961 cgcgcttctc gcgcctgaag gtcatggacg cgattccgat tgtctcggaa ttgctcgccc
     4021 tgggcgtgac gattgtttcc actcaggaag gcgtcttccg gcagggaaac gtcatggacc
     4081 tgattcacct gattatgcgg ctcgacgcgt cgcacaaaga atcttcgctg aagtcggcga
     4141 agattctcga cacgaagaac cttcagcgcg aattgggcgg gtacgtcggc gggaaggcgc
     4201 cttacggctt cgagcttgtt tcggagacga aggagatcac gcgcaacggc cgaatggtca
     4261 atgtcgtcat caacaagctt gcgcactcga ccactcccct taccggaccc ttcgagttcg
     4321 agcccgacgt aatccggtgg tggtggcgtg agatcaagac gcacaaacac cttcccttca
     4381 agccgggcag tcaagccgcc attcacccgg gcagcatcac ggggctttgt aagcgcatgg
     4441 acgctgacgc cgtgccgacc cggggcgaga cgattgggaa gaagaccgct tcaagcgcct
     4501 gggacccggc aaccgttatg cgaatccttc gggacccgcg tattgcgggc ttcgccgctg
     4561 aggtgatcta caagaagaag ccggacggca cgccgaccac gaagattgag ggttaccgca
     4621 ttcagcgcga cccgatcacg ctccggccgg tcgagcttga ttgcggaccg atcatcgagc
     4681 ccgctgagtg gtatgagctt caggcgtggt tggacggcag ggggcgcggc aaggggcttt
     4741 cccgggggca agccattctg tccgccatgg acaagctgta ctgcgagtgt ggcgccgtca
     4801 tgacttcgaa gcgcggggaa gaatcgatca aggactctta ccgctgccgt cgccggaagg
     4861 tggtcgaccc gtccgcacct gggcagcacg aaggcacgtg caacgtcagc atggcggcac
     4921 tcgacaagtt cgttgcggaa cgcatcttca acaagatcag gcacgccgaa ggcgacgaag
     4981 agacgttggc gcttctgtgg gaagccgccc gacgcttcgg caagctcact gaggcgcctg
     5041 agaagagcgg cgaacgggcg aaccttgttg cggagcgcgc cgacgccctg aacgcccttg
     5101 aagagctgta cgaagaccgc gcggcaggcg cgtacgacgg acccgttggc aggaagcact
     5161 tccggaagca acaggcagcg ctgacgctcc ggcagcaagg ggcggaagag cggcttgccg
     5221 aacttgaagc cgccgaagcc ccgaagcttc cccttgacca atggttcccc gaagacgccg
     5281 acgctgaccc gaccggccct aagtcgtggt gggggcgcgc gtcagtagac gacaagcgcg
     5341 tgttcgtcgg gctcttcgta gacaagatcg ttgtcacgaa gtcgactacg ggcagggggc
     5401 agggaacgcc catcgagaag cgcgcttcga tcacgtgggc gaagccgccg accgacgacg
     5461 acgaagacga cgcccaggac ggcacggaag acgtagcggc gtagcgagac acccggatcc
     5521 ctcgaggggc ccaagcttac gcgtgcatgc gacgtcatag ctctctccct atagtgagtc
     5581 gtattataag ctagcttggg atctttgtga aggaacctta cttctgtggt gtgacataat
     5641 tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat
     5701 aatgtgttaa actagctgca tatgcttgct gcttgagagt tttgcttact gagtatgatt
     5761 tatgaaaata ttatacacag gagctagtga ttctaattgt ttgtgtattt tagattcaca
     5821 gtcccaaggc tcatttcagg cccctcagtc ctcacagtct gttcatgatc ataatcagcc
     5881 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     5941 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     6001 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6061 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcgat cctgcattaa
     6121 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg gctggcgtaa tagcgaagag
     6181 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
//
