LOCUS       sequence-244152-        6462 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 6462)
  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 6462)
  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..6462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     996..1015
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1014..1069
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1115..1134
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1220..3037)
                     /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"
     promoter        complement(3044..4255)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3188..3207)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     promoter        complement(4274..4292)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4313..4329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4313..4329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4326..4348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4337..4353
                     /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(4361..4367,4368..4385,4386..4391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4406..4427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4544..4561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4715..5303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4795..4814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5474..6265,6266..6334))
                     /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     6097..6116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6335..6439)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacccatga
      661 ttacgccaag cttgggctgc aggtcgaggg atcttcataa gagaagaggg acagctatga
      721 ctgggagtag tcaggagagg aggaaaaatc tggctagtaa aacatgtaag gaaaatttta
      781 gggatgttaa agaaaaaaat aacacaaaac aaaatataaa aaaaatctaa cctcaagtca
      841 aggcttttct atggaataag gaatggacag cagggggctg tttcatatac tgatgacctc
      901 tttatagcca cctttgttca tggcagccag catatggcat atgttgccaa actctaaacc
      961 aaatactcat tctgatgttt taaatgattt gccctcccat atgtccttcc gagtgagaga
     1021 cacaaaaaat tccaacacac tattgcaatg aaaataaatt tcctttatta gccagaagtc
     1081 agatgctcaa ggggcttcat gatgtcccca taatttttgg cagagggaaa aagatctcag
     1141 tggtatttgt gagccagggc attggccaca ccagccacca ccttctgata ggcagcctgc
     1201 acctgaggag tgcggccgcc taagcagcaa catcttcggt tccgtcttga gcatcgtcct
     1261 catcgtcgtc cgtcgggggc tttgcccatg ttatgctggc ccgcttctct attggggtac
     1321 cttgaccccg accagtagtt gacttggtta ccacaatctt gtccacgaac aagccaacaa
     1381 acactctctt atcatcgacg gacgcccgcc cccaccagct cttaggccct gttggatctg
     1441 catcggcatc ttcggggaac cactggtcca agggcaactt gggggcctct gccgcttcca
     1501 actcagcgag tcgttcctcc gccccctgct ggcgaagagt aagtgcggcc tgttgtttcc
     1561 taaagtgctt tcgcccgact ggaccgtcat aggcgcctgc tgcccggtcc tcataaagtt
     1621 cttccaacgc gttgagtgca tccgctctct ccgctacaag gttagcgcgt tcaccacttt
     1681 tttcaggggc ctccgtcagt ttcccaaagc gccgggctgc ctcccagagc agtgcaagag
     1741 tctcctcgtc tccctcggcg tgtctaatct tattgaatat ccgctcggct acaaacttgt
     1801 cgagcgcagc cattgataca ttgcatgtcc cctcatgttg accaggtgca gaggggtcca
     1861 caactttgcg ccttctgcat cgatatgaat ccttgatgct ttcttctcct cttttggagg
     1921 tcattaccgc accacactcg caatacagtt tatccatggc ggacagtata gcctggcctc
     1981 ggctcaggcc cttcccgcgc cccctaccgt cgagccacgc ctggagttca taccattcag
     2041 cgggctcaat tatcggaccg caatccagct caacaggcct gagagtaatc ggatctcgct
     2101 gaatccggta cccttctatt ttggtggtag gtgtgccatc tggctttttt ttatatataa
     2161 cctcggctgc aaagcccgcg atgcgtgggt ctctaagtat gcgcataacc gtggccggat
     2221 cccaggctga tgaagccgtc tttttgccga tcgtttcccc tcgggtaggc accgcatcag
     2281 catccatccg tttgcacaaa cctgtgatag aaccaggatg aattgcggcc tgtgaccccg
     2341 gcttaaacgg aaggtgctta tgcgtcttga tctcccgcca ccaccatcta atcacgtcgg
     2401 gttcaaactc gaaggggccg gtcagtgggg ttgtgctgtg agccagtttg tttattacaa
     2461 cattaaccat gcgcccattc ctggtaatct ccttcgtttc gctcaccaat tcgaaaccgt
     2521 atggggcctt tccaccaaca taaccaccca actcgcgctg caggttctta gtatcgagta
     2581 tcttcgcgga cttgagggat gattccttgt gggaagcgtc caggcgcata ataagatgta
     2641 taagatccat tacgttcccc tgcctgaaga caccttcttg ggtgctgacg atggtcactc
     2701 cgagagcgag aagctctgag acgattggaa tagcgtccat caccttgagc ctggagaagc
     2761 gtgacacgtc atatactatg atcatgttga gcctgccggc cctacattca ttcaggatgc
     2821 gctcaaactc aggtcgttct gccgtgccga atgcggaggt tcctggagcc tcggaaaaat
     2881 gtcctacaaa tctaaaccta cctccatctc tttctacttc ccgctggagg tctgccgctt
     2941 tatcttcatt ggcggatcgc tgggtcgccg ggcttgcggc actgctgttc tctcgctcgc
     3001 gggactgccg atcatacgct cctgcgtatg tgtccatgaa ttcgtctaac aaaaaagcca
     3061 aaaacggcca gaatttagcg gacaatttac tagtctaaca ctgaaaatta catattgacc
     3121 caaatgatta catttcaaaa ggtgcctaaa aaacttcaca aaacacactc gccaaccccg
     3181 agcgcatagt tcaaaaccgg agcttcagct acttaagaag ataggtacat aaaaccgacc
     3241 aaagaaactg acgcctcact tatccctccc ctcaccagag gtccggcgcc tgtcgattca
     3301 ggagagccta ccctaggccc gaaccctgcg tcctgcgacg gagaaaagcc taccgcacac
     3361 ctaccggcag gtggccccac cctgcattat aagccaacag aacgggtgac gtcacgacac
     3421 gacgaggcgc gcgctcccaa aggtacgggt gcactgccca acggcaccgc ataactgccg
     3481 cccccgcaac cagacgacaa acccgagttc tccagtcagt gacaaacttc acgtcagggt
     3541 ccccagatgg tgccccagcc catctcaccc gaataagagc tttcccgcat tagcgaaggc
     3601 ctcaagacct tgggttcttg ccgcccacca tgccccccac cttgtttcaa cgacctcaca
     3661 gcccgcctca caagcgtctt ccattcaaga ctcgggaaca gccgccattt tgctgcgctc
     3721 cccccaaccc ccagttcagg gcaaccttgc tcgcggaccc agactacagc ccttggcggt
     3781 ctctccacac gcttccgtcc caccgagcgg cccggcggcc acgaaagccc cggccagccc
     3841 agcagcccgc tactcaccaa gtgacgatca cagcgatcca caaacaagaa ccgcgaccca
     3901 aatcccggct gcgacggaac tagctgtgcc acacccggcg cgtccttata taatcatcgg
     3961 cgttcaccgc cccacggaga tccctccgca gaatcgccga gaagggacta cttttcctcg
     4021 cctgttccgc tctctggaaa gaaaaccagt gccctagagt cacccaagtc ccgtcctaaa
     4081 atgtccttct gctgatactg gggttctaag gccgagtctt atgagcagcg ggccgctgtc
     4141 ctgagcgtcc gggcggaagg atcaggacgc tcgctgcgcc cttcgtctga cgtggcagcg
     4201 ctcgccgtga ggaggggggc gcccgcggga ggcgccaaaa cccggcgcgg aggccgagct
     4261 ccagcttttg ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc
     4321 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4381 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4441 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4501 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4561 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4621 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4681 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4741 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4801 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4861 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4921 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4981 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5041 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5101 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5161 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5221 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5281 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5341 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5401 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5461 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5521 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5581 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5641 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5701 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5761 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5821 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5881 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5941 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6001 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6061 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6121 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6181 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6241 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6301 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6361 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6421 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc ac
//
