LOCUS       sequence-434496-        5907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the overexpression of phiC31 integrase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5907)
  AUTHORS   Hong J, Sohn KC, Park HW, Jeon H, Ju E, Lee JG, Lee JS, Rho J, Hur 
            GM, Ro H
  TITLE     All-in-one IQ toggle switches with high versatilities for 
            fine-tuning of transgene expression in mammalian cells and tissues.
  JOURNAL   Mol Ther Methods Clin Dev. 2024 Feb 2;32(1):101202. doi: 
            10.1016/j.omtm.2024.101202. eCollection 2024 Mar 14.
  PUBMED    38374964
REFERENCE   2  (bases 1 to 5907)
  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: "Mol Ther 
            Methods Clin Dev."; date: "2024-02-2"; volume: "32(1):101202. doi"; 
            pages: " 10.1016/j.omtm.2024.101202. eCollection 2024 Mar 14"
COMMENT     Sequence Label: pCS2+ phiC31 integrase
FEATURES             Location/Qualifiers
     source          1..5907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      99..108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             105..1922
                     /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"
     polyA_signal    1959..2093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1983..2002)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2037..2056
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2206..2226)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2206..2224)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2245..2261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2245..2261)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2258..2280)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2269..2285
                     /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(join(2293..2299,2300..2317,2318..2323))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2338..2359
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2476..2493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2647..3235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2727..2746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3406..4197,4198..4266))
                     /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     4029..4048
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4267..4371)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4397..4852)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4534..4555)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4746..4765
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4923..5907
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattgc caccatggac acgtacgcgg
      121 gtgcttacga ccgtcagtcg cgcgagcgcg aaaattcgag cgcagcaagc ccagcgacac
      181 agcgtagcgc caacgaagac aaggcggccg accttcagcg cgaagtcgag cgcgacgggg
      241 gccggttcag gttcgtcggg catttcagcg aagcgccggg cacgtcggcg ttcgggacgg
      301 cggagcgccc ggagttcgaa cgcatcctga acgaatgccg cgccgggcgg ctcaacatga
      361 tcattgtcta tgacgtgtcg cgcttctcgc gcctgaaggt catggacgcg attccgattg
      421 tctcggaatt gctcgccctg ggcgtgacga ttgtttccac tcaggaaggc gtcttccggc
      481 agggaaacgt catggacctg attcacctga ttatgcggct cgacgcgtcg cacaaagaat
      541 cttcgctgaa gtcggcgaag attctcgaca cgaagaacct tcagcgcgaa ttgggcgggt
      601 acgtcggcgg gaaggcgcct tacggcttcg agcttgtttc ggagacgaag gagatcacgc
      661 gcaacggccg aatggtcaat gtcgtcatca acaagcttgc gcactcgacc actcccctta
      721 ccggaccctt cgagttcgag cccgacgtaa tccggtggtg gtggcgtgag atcaagacgc
      781 acaaacacct tcccttcaag ccgggcagtc aagccgccat tcacccgggc agcatcacgg
      841 ggctttgtaa gcgcatggac gctgacgccg tgccgacccg gggcgagacg attgggaaga
      901 agaccgcttc aagcgcctgg gacccggcaa ccgttatgcg aatccttcgg gacccgcgta
      961 ttgcgggctt cgccgctgag gtgatctaca agaagaagcc ggacggcacg ccgaccacga
     1021 agattgaggg ttaccgcatt cagcgcgacc cgatcacgct ccggccggtc gagcttgatt
     1081 gcggaccgat catcgagccc gctgagtggt atgagcttca ggcgtggttg gacggcaggg
     1141 ggcgcggcaa ggggctttcc cgggggcaag ccattctgtc cgccatggac aagctgtact
     1201 gcgagtgtgg cgccgtcatg acttcgaagc gcggggaaga atcgatcaag gactcttacc
     1261 gctgccgtcg ccggaaggtg gtcgacccgt ccgcacctgg gcagcacgaa ggcacgtgca
     1321 acgtcagcat ggcggcactc gacaagttcg ttgcggaacg catcttcaac aagatcaggc
     1381 acgccgaagg cgacgaagag acgttggcgc ttctgtggga agccgcccga cgcttcggca
     1441 agctcactga ggcgcctgag aagagcggcg aacgggcgaa ccttgttgcg gagcgcgccg
     1501 acgccctgaa cgcccttgaa gagctgtacg aagaccgcgc ggcaggcgcg tacgacggac
     1561 ccgttggcag gaagcacttc cggaagcaac aggcagcgct gacgctccgg cagcaagggg
     1621 cggaagagcg gcttgccgaa cttgaagccg ccgaagcccc gaagcttccc cttgaccaat
     1681 ggttccccga agacgccgac gctgacccga ccggccctaa gtcgtggtgg gggcgcgcgt
     1741 cagtagacga caagcgcgtg ttcgtcgggc tcttcgtaga caagatcgtt gtcacgaagt
     1801 cgactacggg cagggggcag ggaacgccca tcgagaagcg cgcttcgatc acgtgggcga
     1861 agccgccgac cgacgacgac gaagacgacg cccaggacgg cacggaagac gtagcggcgt
     1921 agtcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     1981 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2041 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2101 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     2161 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     2221 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2281 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     2341 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2401 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2461 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2521 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2581 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2641 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2701 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2761 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2821 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2881 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2941 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3001 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3061 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3121 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3181 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3241 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3301 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3361 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3421 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3481 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3541 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3601 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3661 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3721 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3781 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3841 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3901 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3961 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4021 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4081 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4141 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4201 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4261 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4321 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4381 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4441 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4501 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4561 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4621 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4681 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4741 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4801 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     4861 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4921 cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc
     4981 tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc
     5041 tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggacc
     5101 tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc
     5161 ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat
     5221 tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc
     5281 tattggccag gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt
     5341 tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct
     5401 tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt
     5461 cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca
     5521 attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat
     5581 tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt
     5641 caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa
     5701 gtacattact attggaagta cgccagggta cattggcagt actcccattg acgtcaatgg
     5761 cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca
     5821 atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc
     5881 tatataagca atgctcgttt agggaac
//
