LOCUS       sequence-201482-        7524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Carries phiC31 integrase attB site flanked by cHS4 insulators..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7524)
  AUTHORS   Zhao C, Farruggio AP, Bjornson CR, Chavez CL, Geisinger JM, Neal TL,
            Karow M, Calos MP
  TITLE     Recombinase-mediated reprogramming and dystrophin gene addition in 
            mdx mouse induced pluripotent stem cells.
  JOURNAL   PLoS One. 2014 Apr 29;9(4):e96279. doi: 
            10.1371/journal.pone.0096279. eCollection 2014.
  PUBMED    24781921
REFERENCE   2  (bases 1 to 7524)
  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: "PLoS One.";
            date: "2014-04-29"; volume: "9(4):e96279. doi"; pages: " 
            10.1371/journal.pone.0096279. eCollection 2014"
COMMENT     Sequence Label: pC-HBH
FEATURES             Location/Qualifiers
     source          1..7524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        359..636
                     /label=chicken beta-actin promoter
     intron          637..1652
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1576..1598
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1660..1679
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(1746..1763)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1752..1976
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(1985..2018)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3293..3326
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    3525..3594
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    3610..3643
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        4970..5074
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5076..5433
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5076..5267
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5099..5119)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5284..5419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5468..6262
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5468..6262
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5522..5541)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6132..6151
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6450..6469)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6494..6541
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6870..7458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7359..7378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
      361 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
      421 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
      481 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      541 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      601 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      661 cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
      721 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
      781 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
      841 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
      901 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
      961 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     1021 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     1081 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     1141 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     1201 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     1261 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     1321 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     1381 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     1441 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     1501 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     1561 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     1621 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     1681 tgtctcatca ttttggcaaa gaattccgtt cggacgaaag ttgttgaaac gcgcgcgact
     1741 cgagccctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     1801 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     1861 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     1921 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggggcg
     1981 cgccataact tcgtatagca tacattatac gaagttatag atctcaggtt tcagtctcag
     2041 tcaaagggct agagccccat cctcactgac tccgtcctgg agttggatga gagataatgg
     2101 ccttacgttg tgccagggga gggtcgggct ggatttagca agatttacct tctccaaaga
     2161 gcggtgctgc agtggcacag ctgcccacgg aggtgggggg gtcaccgtcc ctggaggtga
     2221 tgaagaactg tggggatgtg gcactgaggg acatggccag tgggcacggt gggtgggttg
     2281 gggttggtct tggggatctt ggagggcttt tccagccttc atgatttgac gattgtatga
     2341 acatctacat ggcaattctc cagctgcctg tcccagtcct actgacccag ctgtatctct
     2401 ccaggcaagc tcttccaccc cttctgcttg catccagaca ccatcaaaca tgcaggctca
     2461 gacacaggga ccagcagtgt ctgtggcctt tttgtgctcc tctccatgct gggttttaac
     2521 ttgctctttg tccttctatc ctatcttctt atccttaagg ctgttctgaa cgctgtgact
     2581 tggagagtgt cccagagccc tcaacacctg catgtcccac gtccatgctg tcctgcactt
     2641 ccttatcccc aagatctgcc tctccgtgat gcactgaatt ggcaaacatg tgtcacccca
     2701 gaccaacaat gtcacagcaa actccccctt gataggacaa gggggaatgg ctttacactg
     2761 agacagggga ggtttgggtt ggatatgagg aggcagtttt tcccccagag ggtggtgacg
     2821 cactgaacag gttgcccaag gaggctgtgg atgccccatc cctgcaggca ttcaaggcca
     2881 ggctggatgt ggctctgggc agcctgggct gctggttgat gaccctgcac atagcagggg
     2941 gttggatctg gatgagcact gtgctccttt gcaacccagg ccgttctatg attctgtcat
     3001 tctaaatctc tctttcagcc taaagctttt tccccgtatc cccccaggtg tctgcaggct
     3061 caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg
     3121 gctgtccccg cacgctgccg gctcggggat gcggggggag cgccggaccg gagcggagcc
     3181 ccgggcggct cgctgctgcc ccctagcggg ggagggacgt aattacatcc ctgggggctt
     3241 tggggggggg ctgtccctct agcttccctg tgtttgttct ccacctaagc ttgaagttcc
     3301 tattctctag aaagtatagg aacttcggct gtcgacatgc ccgccgtgac cgtcgagaac
     3361 ccgctgacgc tgccccgcgt atccgcaccc gccgacgccg tcgcacgtcc cgtgctcacc
     3421 gtgaccaccg cgcccagcgg tttcgagggc gagggcttcc cggtgcgccg cgcgttcgcc
     3481 gggatcaact accgccacct cgacccgttc atcatgatgg accagatggg tgaggtggag
     3541 tacgcgcccg gggagcccaa gggcacgccc tggcacccgc accgcggctt cgagaccgtg
     3601 acctacatcg aagttcctat tctctagaaa gtataggaac ttcactagtt tgagggtgtg
     3661 atgtttggag aggctagagg gacagccccc ccccaaagcc cccagggatg taattacgtc
     3721 cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc cggcgctccc
     3781 cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag gtggcacggg
     3841 atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct ggggggatac
     3901 ggggaaaaag ctttaggctg aaagagagat ttagaatgac agaatcatag aacggcctgg
     3961 gttgcaaagg agcacagtgc tcatccagat ccaaccccct gctatgtgca gggtcatcaa
     4021 ccagcagccc aggctgccca gagccacatc cagcctggcc ttgaatgcct gcagggatgg
     4081 ggcatccaca gcctccttgg gcaacctgtt cagtgcgtca ccaccctctg ggggaaaaac
     4141 tgcctcctca tatccaaccc aaacctcccc tgtctcagtg taaagccatt cccccttgtc
     4201 ctatcaaggg ggagtttgct gtgacattgt tggtctgggg tgacacatgt ttgccaattc
     4261 agtgcatcac ggagaggcag atcttgggga taaggaagtg caggacagca tggacgtggg
     4321 acatgcaggt gttgagggct ctgggacact ctccaagtca cagcgttcag aacagcctta
     4381 aggataagaa gataggatag aaggacaaag agcaagttaa aacccagcat ggagaggagc
     4441 acaaaaaggc cacagacact gctggtccct gtgtctgagc ctgcatgttt gatggtgtct
     4501 ggatgcaagc agaaggggtg gaagagcttg cctggagaga tacagctggg tcagtaggac
     4561 tgggacaggc agctggagaa ttgccatgta gatgttcata caatcgtcaa atcatgaagg
     4621 ctggaaaagc cctccaagat ccccaagacc aaccccaacc cacccaccgt gcccactggc
     4681 catgtccctc agtgccacat ccccacagtt cttcatcacc tccagggacg gtgacccccc
     4741 cacctccgtg ggcagctgtg ccactgcagc accgctcttt ggagaaggta aatcttgcta
     4801 aatccagccc gaccctcccc tggcacaacg taaggccatt atctctcatc caactccagg
     4861 acggagtcag tgaggatggg gctctaggac aagagacaca aagtccccaa ggcggccgct
     4921 taatttgttg cgatcgctcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4981 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5041 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     5101 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5161 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5221 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5281 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5341 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5401 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     5461 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5521 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5581 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5641 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5701 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5761 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5821 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5881 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5941 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     6001 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     6061 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6121 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6181 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6241 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6301 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6361 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6421 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     6481 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     6541 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     6601 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     6661 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     6721 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6781 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6841 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6901 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6961 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7021 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7081 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7141 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7201 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7261 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7321 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7381 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7441 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7501 tggccttttg ctctcctgca ggat
//
