LOCUS       sequence-250824-        6463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes H. sapiens codon optimized Integrase phiC31..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6463)
  AUTHORS   Gomide MS, Sales TT, Barros LRC, Limia CG, de Oliveira MA, 
            Florentino LH, Barros LMG, Robledo ML, Jose GPC, Almeida MSM, Lima 
            RN, Rehen SK, Lacorte C, Melo EO, Murad AM, Bonamino MH, Coelho CM, 
            Rech E
  TITLE     Genetic switches designed for eukaryotic cells and controlled by 
            serine integrases.
  JOURNAL   Commun Biol. 2020 May 22;3(1):255. doi: 10.1038/s42003-020-0971-8.
  PUBMED    32444777
REFERENCE   2  (bases 1 to 6463)
  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: "Commun 
            Biol."; date: "2020-05-22"; volume: "3(1):255. doi"; pages: " 
            10.1038/s42003-020-0971-8"
COMMENT     Sequence Label: INCTbiosyn-pUB-HspINTphiC31
FEATURES             Location/Qualifiers
     source          1..6463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    227..248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(263..268,269..286,287..293)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    301..317
                     /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     306..328
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     325..341
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     325..341
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        362..380
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        399..1610
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1447..1466
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1617..3434
                     /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(3520..3539)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3585..3640
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3639..3658)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4012..4031)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4013..4031)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4038..4055)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4038..4054)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4047..4069)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4196..4651)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4333..4354)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4545..4564
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4678..4782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4783..4851,4852..5643)
                     /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(5001..5020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5814..6402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6303..6322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
       61 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
      121 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
      181 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      241 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      301 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg
      361 aaattaaccc tcactaaagg gaacaaaagc tggagctcgg cctccgcgcc gggttttggc
      421 gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag
      481 cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
      541 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
      601 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
      661 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
      721 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg
      781 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
      841 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg
      901 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc
      961 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg
     1021 gcatggtggg cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta
     1081 ttcgggtgag atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     1141 ggagaactcg ggtttgtcgt ctggttgcgg gggcggcagt tatgcggtgc cgttgggcag
     1201 tgcacccgta cctttgggag cgcgcgcctc gtcgtgtcgt gacgtcaccc gttctgttgg
     1261 cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca
     1321 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
     1381 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
     1441 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
     1501 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
     1561 ctagtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac gaattcatgg
     1621 acacatacgc aggagcgtat gatcggcagt cccgcgagcg agagaacagc agtgccgcaa
     1681 gcccggcgac ccagcgatcc gccaatgaag ataaagcggc agacctccag cgggaagtag
     1741 aaagagatgg aggtaggttt agatttgtag gacatttttc cgaggctcca ggaacctccg
     1801 cattcggcac ggcagaacga cctgagtttg agcgcatcct gaatgaatgt agggccggca
     1861 ggctcaacat gatcatagta tatgacgtgt cacgcttctc caggctcaag gtgatggacg
     1921 ctattccaat cgtctcagag cttctcgctc tcggagtgac catcgtcagc acccaagaag
     1981 gtgtcttcag gcaggggaac gtaatggatc ttatacatct tattatgcgc ctggacgctt
     2041 cccacaagga atcatccctc aagtccgcga agatactcga tactaagaac ctgcagcgcg
     2101 agttgggtgg ttatgttggt ggaaaggccc catacggttt cgaattggtg agcgaaacga
     2161 aggagattac caggaatggg cgcatggtta atgttgtaat aaacaaactg gctcacagca
     2221 caaccccact gaccggcccc ttcgagtttg aacccgacgt gattagatgg tggtggcggg
     2281 agatcaagac gcataagcac cttccgttta agccggggtc acaggccgca attcatcctg
     2341 gttctatcac aggtttgtgc aaacggatgg atgctgatgc ggtgcctacc cgaggggaaa
     2401 cgatcggcaa aaagacggct tcatcagcct gggatccggc cacggttatg cgcatactta
     2461 gagacccacg catcgcgggc tttgcagccg aggttatata taaaaaaaag ccagatggca
     2521 cacctaccac caaaatagaa gggtaccgga ttcagcgaga tccgattact ctcaggcctg
     2581 ttgagctgga ttgcggtccg ataattgagc ccgctgaatg gtatgaactc caggcgtggc
     2641 tcgacggtag ggggcgcggg aagggcctga gccgaggcca ggctatactg tccgccatgg
     2701 ataaactgta ttgcgagtgt ggtgcggtaa tgacctccaa aagaggagaa gaaagcatca
     2761 aggattcata tcgatgcaga aggcgcaaag ttgtggaccc ctctgcacct ggtcaacatg
     2821 aggggacatg caatgtatca atggctgcgc tcgacaagtt tgtagccgag cggatattca
     2881 ataagattag acacgccgag ggagacgagg agactcttgc actgctctgg gaggcagccc
     2941 ggcgctttgg gaaactgacg gaggcccctg aaaaaagtgg tgaacgcgct aaccttgtag
     3001 cggagagagc ggatgcactc aacgcgttgg aagaacttta tgaggaccgg gcagcaggcg
     3061 cctatgacgg tccagtcggg cgaaagcact ttaggaaaca acaggccgca cttactcttc
     3121 gccagcaggg ggcggaggaa cgactcgctg agttggaagc ggcagaggcc cccaagttgc
     3181 ccttggacca gtggttcccc gaagatgccg atgcagatcc aacagggcct aagagctggt
     3241 gggggcgggc gtccgtcgat gataagagag tgtttgttgg cttgttcgtg gacaagattg
     3301 tggtaaccaa gtcaactact ggtcggggtc aaggtacccc aatagagaag cgggccagca
     3361 taacatgggc aaagcccccg acggacgacg atgaggacga tgctcaagac ggaaccgaag
     3421 atgttgctgc ttaggcggcc gcactcctca ggtgcaggct gcctatcaga aggtggtggc
     3481 tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa
     3541 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     3601 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     3661 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccat
     3721 atgctggctg ccatgaacaa aggtggctat aaagaggtca tcagtatatg aaacagcccc
     3781 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     3841 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     3901 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgaa
     3961 gatccctcga cctgcagccc aagcttggcg taatcatggg tacccaattc gccctatagt
     4021 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4081 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4141 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcaaatt
     4201 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4261 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4321 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4381 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4441 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4501 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4561 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4621 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     4681 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4741 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4801 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4861 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4921 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4981 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5041 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5101 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5161 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5221 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5281 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5341 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5401 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5461 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5521 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5581 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5641 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5701 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5761 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5821 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5881 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5941 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6001 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6061 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6121 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6181 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6241 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6301 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6361 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6421 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
