LOCUS       sequence-219880-        6228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6228)
  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 6228)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2000 May 23. 97(11):5995-6000."
COMMENT     Sequence Label: pCMVInt
FEATURES             Location/Qualifiers
     source          1..6228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(189..293)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     361..379
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(406..411,412..429,430..436)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    444..460
                     /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     449..471
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     enhancer        490..793
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        794..997
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     948..968
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     994..1018
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1113..1131
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1113..1130
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1224..3041
                     /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(3104..3123)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3105..3123)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3510..3644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3547..3566)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3601..3620
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3770..4224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3857..3876)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4066..4087
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(4511..4579,4580..5371)
                     /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(4729..4748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    5473..5506
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      5584..6172
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6073..6092
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 agcttgcaat tcgcgctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      301 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
      361 aataggcgta tcacgaggcc ctttcactca ttaggcaccc caggctttac actttatgct
      421 tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcat
      481 cgtgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      541 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      601 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      661 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      721 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      781 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      841 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      901 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      961 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     1021 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     1081 ccggactcta gcctaggctt ttgcaaaaag ctatttaggt gacactatag aaggtacgcc
     1141 tgcaggtacc ggtccggaat tcccgggtcg acgagctcac tagtcgtagg gtcgccgaca
     1201 tgacacaagg ggttgtgacc ggggtggaca cgtacgcggg tgcttacgac cgtcagtcgc
     1261 gcgagcgcga aaattcgagc gcagcaagcc cagcgacaca gcgtagcgcc aacgaagaca
     1321 aggcggccga ccttcagcgc gaagtcgagc gcgacggggg ccggttcagg ttcgtcgggc
     1381 atttcagcga agcgccgggc acgtcggcgt tcgggacggc ggagcgcccg gagttcgaac
     1441 gcatcctgaa cgaatgccgc gccgggcggc tcaacatgat cattgtctat gacgtgtcgc
     1501 gcttctcgcg cctgaaggtc atggacgcga ttccgattgt ctcggaattg ctcgccctgg
     1561 gcgtgacgat tgtttccact caggaaggcg tcttccggca gggaaacgtc atggacctga
     1621 ttcacctgat tatgcggctc gacgcgtcgc acaaagaatc ttcgctgaag tcggcgaaga
     1681 ttctcgacac gaagaacctt cagcgcgaat tgggcgggta cgtcggcggg aaggcgcctt
     1741 acggcttcga gcttgtttcg gagacgaagg agatcacgcg caacggccga atggtcaatg
     1801 tcgtcatcaa caagcttgcg cactcgacca ctccccttac cggacccttc gagttcgagc
     1861 ccgacgtaat ccggtggtgg tggcgtgaga tcaagacgca caaacacctt cccttcaagc
     1921 cgggcagtca agccgccatt cacccgggca gcatcacggg gctttgtaag cgcatggacg
     1981 ctgacgccgt gccgacccgg ggcgagacga ttgggaagaa gaccgcttca agcgcctggg
     2041 acccggcaac cgttatgcga atccttcggg acccgcgtat tgcgggcttc gccgctgagg
     2101 tgatctacaa gaagaagccg gacggcacgc cgaccacgaa gattgagggt taccgcattc
     2161 agcgcgaccc gatcacgctc cggccggtcg agcttgattg cggaccgatc atcgagcccg
     2221 ctgagtggta tgagcttcag gcgtggttgg acggcagggg gcgcggcaag gggctttccc
     2281 gggggcaagc cattctgtcc gccatggaca agctgtactg cgagtgtggc gccgtcatga
     2341 cttcgaagcg cggggaagaa tcgatcaagg actcttaccg ctgccgtcgc cggaaggtgg
     2401 tcgacccgtc cgcacctggg cagcacgaag gcacgtgcaa cgtcagcatg gcggcactcg
     2461 acaagttcgt tgcggaacgc atcttcaaca agatcaggca cgccgaaggc gacgaagaga
     2521 cgttggcgct tctgtgggaa gccgcccgac gcttcggcaa gctcactgag gcgcctgaga
     2581 agagcggcga acgggcgaac cttgttgcgg agcgcgccga cgccctgaac gcccttgaag
     2641 agctgtacga agaccgcgcg gcaggcgcgt acgacggacc cgttggcagg aagcacttcc
     2701 ggaagcaaca ggcagcgctg acgctccggc agcaaggggc ggaagagcgg cttgccgaac
     2761 ttgaagccgc cgaagccccg aagcttcccc ttgaccaatg gttccccgaa gacgccgacg
     2821 ctgacccgac cggccctaag tcgtggtggg ggcgcgcgtc agtagacgac aagcgcgtgt
     2881 tcgtcgggct cttcgtagac aagatcgttg tcacgaagtc gactacgggc agggggcagg
     2941 gaacgcccat cgagaagcgc gcttcgatca cgtgggcgaa gccgccgacc gacgacgacg
     3001 aagacgacgc ccaggacggc acggaagacg tagcggcgta gcgagacacc cggatccctc
     3061 gaggggccca agcttacgcg tgcatgcgac gtcatagctc tctccctata gtgagtcgta
     3121 ttataagcta gcttgggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg
     3181 acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat
     3241 gtgttaaact agctgcatat gcttgctgct tgagagtttt gcttactgag tatgatttat
     3301 gaaaatatta tacacaggag ctagtgattc taattgtttg tgtattttag attcacagtc
     3361 ccaaggctca tttcaggccc ctcagtcctc acagtctgtt catgatcata atcagccata
     3421 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3481 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3541 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3601 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcgatcct gcattaatga
     3661 atcggccaac gcgcggggag aggcggtttg cgtattggct ggcgtaatag cgaagaggcc
     3721 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     3781 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3841 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3901 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3961 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4021 tagacggttt ttcgcccttt gacgttggag tccacgttct taatagtgga ctcttgttcc
     4081 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4141 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4201 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4261 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgccag gtcttggact
     4321 ggtgagaacg gcttgctcgg cagcttcgat gtgtgctgga gggagaataa aggtctaaga
     4381 tgtgcgatag agggaagtcg cattgaatta tgtgctgtgt agggatcgct ggtatcaaat
     4441 atgtgtgccc acccctggca tgagacaata accctgataa atgcttcaat aatattgaaa
     4501 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4561 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4621 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4681 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4741 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4801 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4861 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4921 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4981 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5041 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5101 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5161 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5221 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5281 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5341 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5401 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5461 taatctcatg ccataacttc gtataatgta tgctatacga agttatggca tgaccaaaat
     5521 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5581 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5641 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5701 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5761 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5821 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5881 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5941 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6001 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6061 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6121 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6181 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctca
//
