LOCUS       sequence-261191-        6204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian phiC31 integrase expression vector. For use in 
            applications where pseudosite integration is not desired..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6204)
  AUTHORS   Zhu F, Gamboa M, Farruggio AP, Hippenmeyer S, Tasic B, Schule B, 
            Chen-Tsai Y, Calos MP
  TITLE     DICE, an efficient system for iterative genomic editing in human 
            pluripotent stem cells.
  JOURNAL   Nucleic Acids Res. 2013 Dec 4.
  PUBMED    24304893
REFERENCE   2  (bases 1 to 6204)
  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: "Nucleic 
            Acids Res. 2013 Dec 4."
COMMENT     Sequence Label: pCS-kI
FEATURES             Location/Qualifiers
     source          1..6204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /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     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(743..811,812..1603)
                     /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(961..980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    1705..1738
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      1816..2404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2305..2324
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2558..2575
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(2649..2753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2821..2839
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2866..2871,2872..2889,2890..2896)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2904..2920
                     /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     2909..2931
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     enhancer        2950..3253
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3254..3457
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3408..3428
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3454..3478
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3573..3591
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3573..3590
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      3654..3663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3660..5477
                     /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(5540..5559)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5541..5559)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5946..6080
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5983..6002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6037..6056
                     /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 cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgcc
      541 aggtcttgga ctggtgagaa cggcttgctc ggcagcttcg atgtgtgctg gagggagaat
      601 aaaggtctaa gatgtgcgat agagggaagt cgcattgaat tatgtgctgt gtagggatcg
      661 ctggtatcaa atatgtgtgc ccacccctgg catgagacaa taaccctgat aaatgcttca
      721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      961 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1261 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1441 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1681 gatccttttt gataatctca tgccataact tcgtataatg tatgctatac gaagttatgg
     1741 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1801 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1861 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1921 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1981 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2041 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2101 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2161 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2221 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2281 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2341 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2401 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2461 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2521 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2581 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2641 agcttgcaat tcgcgctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     2701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2761 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     2821 aataggcgta tcacgaggcc ctttcactca ttaggcaccc caggctttac actttatgct
     2881 tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcat
     2941 cgtgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     3001 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     3061 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     3121 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     3181 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     3241 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     3301 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     3361 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     3421 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     3481 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     3541 ccggactcta gcctaggctt ttgcaaaaag ctatttaggt gacactatag aaggtacgcc
     3601 tgcaggtacc ggtccggaat tcccgggtcg acgagctcac tagtcgtagg gtcgccacca
     3661 tggacacgta cgcgggtgct tacgaccgtc agtcgcgcga gcgcgaaaat tcgagcgcag
     3721 caagcccagc gacacagcgt agcgccaacg aagacaaggc ggccgacctt cagcgcgaag
     3781 tcgagcgcga cgggggccgg ttcaggttcg tcgggcattt cagcgaagcg ccgggcacgt
     3841 cggcgttcgg gacggcggag cgcccggagt tcgaacgcat cctgaacgaa tgccgcgccg
     3901 ggcggctcaa catgatcatt gtctatgacg tgtcgcgctt ctcgcgcctg aaggtcatgg
     3961 acgcgattcc gattgtctcg gaattgctcg ccctgggcgt gacgattgtt tccactcagg
     4021 aaggcgtctt ccggcaggga aacgtcatgg acctgattca cctgattatg cggctcgacg
     4081 cgtcgcacaa agaatcttcg ctgaagtcgg cgaagattct cgacacgaag aaccttcagc
     4141 gcgaattggg cgggtacgtc ggcgggaagg cgccttacgg cttcgagctt gtttcggaga
     4201 cgaaggagat cacgcgcaac ggccgaatgg tcaatgtcgt catcaacaag cttgcgcact
     4261 cgaccactcc ccttaccgga cccttcgagt tcgagcccga cgtaatccgg tggtggtggc
     4321 gtgagatcaa gacgcacaaa caccttccct tcaagccggg cagtcaagcc gccattcacc
     4381 cgggcagcat cacggggctt tgtaagcgca tggacgctga cgccgtgccg acccggggcg
     4441 agacgattgg gaagaagacc gcttcaagcg cctgggaccc ggcaaccgtt atgcgaatcc
     4501 ttcgggaccc gcgtattgcg ggcttcgccg ctgaggtgat ctacaagaag aagccggacg
     4561 gcacgccgac cacgaagatt gagggttacc gcattcagcg cgacccgatc acgctccggc
     4621 cggtcgagct tgattgcgga ccgatcatcg agcccgctga gtggtatgag cttcaggcgt
     4681 ggttggacgg cagggggcgc ggcaaggggc tttcccgggg gcaagccatt ctgtccgcca
     4741 tggacaagct gtactgcgag tgtggcgccg tcatgacttc gaagcgcggg gaagaatcga
     4801 tcaaggactc ttaccgctgc cgtcgccgga aggtggtcga cccgtccgca cctgggcagc
     4861 acgaaggcac gtgcaacgtc agcatggcgg cactcgacaa gttcgttgcg gaacgcatct
     4921 tcaacaagat caggcacgcc gaaggcgacg aagagacgtt ggcgcttctg tgggaagccg
     4981 cccgacgctt cggcaagctc actgaggcgc ctgagaagag cggcgaacgg gcgaaccttg
     5041 ttgcggagcg cgccgacgcc ctgaacgccc ttgaagagct gtacgaagac cgcgcggcag
     5101 gcgcgtacga cggacccgtt ggcaggaagc acttccggaa gcaacaggca gcgctgacgc
     5161 tccggcagca aggggcggaa gagcggcttg ccgaacttga agccgccgaa gccccgaagc
     5221 ttccccttga ccaatggttc cccgaagacg ccgacgctga cccgaccggc cctaagtcgt
     5281 ggtgggggcg cgcgtcagta gacgacaagc gcgtgttcgt cgggctcttc gtagacaaga
     5341 tcgttgtcac gaagtcgact acgggcaggg ggcagggaac gcccatcgag aagcgcgctt
     5401 cgatcacgtg ggcgaagccg ccgaccgacg acgacgaaga cgacgcccag gacggcacgg
     5461 aagacgtagc ggcgtagcga gacacccgga tccctcgagg ggcccaagct tacgcgtgca
     5521 tgcgacgtca tagctctctc cctatagtga gtcgtattat aagctagctt gggatctttg
     5581 tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa
     5641 gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactagct gcatatgctt
     5701 gctgcttgag agttttgctt actgagtatg atttatgaaa atattataca caggagctag
     5761 tgattctaat tgtttgtgta ttttagattc acagtcccaa ggctcatttc aggcccctca
     5821 gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag gttttacttg
     5881 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     5941 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6001 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6061 tatcttatca tgtctggatc gatcctgcat taatgaatcg gccaacgcgc ggggagaggc
     6121 ggtttgcgta ttggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     6181 ttgcgcagcc tgaatggcga atgg
//
