LOCUS       sequence-198846-        7637 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A piggyBac transposon based dual directional gene trap vector, in 
            which, the mutagens are two non-selectable gene-trap cassettes in 
            opposite orientations..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7637)
  AUTHORS   Liu G, Wang X, Liu Y, Zhang M, Cai T, Shen Z, Jia Y, Huang Y
  TITLE     Arrayed mutant haploid embryonic stem cell libraries facilitate 
            phenotype-driven genetic screens.
  JOURNAL   Nucleic Acids Res. 2017 Sep 28. doi: 10.1093/nar/gkx857.
  PUBMED    29036617
REFERENCE   2  (bases 1 to 7637)
  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. 2017 Sep 28. doi: 10.1093/nar/gkx857."
COMMENT     Sequence Label: PBDGTV
FEATURES             Location/Qualifiers
     source          1..7637
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     52..71
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      196..205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             202..801
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     701..721
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             808..1803
                     /codon_start=1
                     /product="truncated herpes simplex virus thymidine kinase 
                     (Salomon et al., 1995)"
                     /label=Delta-TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(1825..1842)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1831..2055
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3154..3216
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3299..3318)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3300..3318)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3325..3342)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3325..3341)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3334..3356)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3483..3938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3570..3589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3780..3801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3964..4068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4069..4137,4138..4929)
                     /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(4287..4306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5100..5688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5589..5608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5842..5859
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5976..5997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6012..6017,6018..6035,6036..6042)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6050..6066
                     /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     6055..6077
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6074..6090
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6074..6090
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6111..6129
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   6192..6226
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7300..7323)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
       61 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
      121 cctgcagata acttcgtata gcatacatta tacgaagcgg gggggcccgg tacggatcct
      181 cgagcagctg ccaacgccac catggggacc gagtacaagc ccacggtgcg cctcgccacc
      241 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
      301 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
      361 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
      421 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
      481 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
      541 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
      601 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
      661 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
      721 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
      781 atgacccgca agcccggtgc cggatccatg cccacgctac tgcgggttta tatagacggt
      841 cctcacggga tggggaaaac caccaccacg caactgctgg tggccctggg ttcgcgcgac
      901 gatatcgtct acgtacccga gccgatgact tactggcagg tgctgggggc ttccgagaca
      961 atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc ggccggggac
     1021 gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc cgtgaccgac
     1081 gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc gcccccggcc
     1141 ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc ggccgcgcga
     1201 taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct catcccgccg
     1261 accttgcccg gcacaaacat cgtgttgggg gcccttccgg aggacagaca catcgaccgc
     1321 ctggccaaac gccagcgccc cggcgagcgg cttgacctgg ctatgctggc cgcgattcgc
     1381 cgcgtttacg ggctgcttgc caatacggtg cggtatctgc agggcggcgg gtcgtggcgg
     1441 gaggattggg gacagctttc ggggacggcc gtgccgcccc agggtgccga gccccagagc
     1501 aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg ggcccccgag
     1561 ttgctggccc ccaacggcga cctgtacaac gtgtttgcct gggccttgga cgtcttggcc
     1621 aaacgcctcc gtcccatgca cgtctttatc ctggattacg accaatcgcc cgccggctgc
     1681 cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac cacccccggc
     1741 tccataccga cgatctgcga cctggcgcgc acgtttgccc gggagatggg ggaggctaac
     1801 tgagctctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1861 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1921 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1981 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2041 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgaga tccactagtt
     2101 ctagcctcga gcaagtcttt cagtcctata gaaataaaat caggaattaa aaagaagatt
     2161 ctgaatattt atcttttata acaaatactt cttacacatt aaacttttac aggacacatt
     2221 ccttgtctct atgaagggaa ttcgcgccgc cagcctctac acccagtaaa ggatgtgtgg
     2281 atagaaatgt aacaataagc tgggttgtag tagcagcgaa ggggcagata aatctctttg
     2341 agtttgatgc caaccatgtt ttgaaaaaac taaaaacaaa caaaaaaaac cccctaaaaa
     2401 cccaaaaata taaataaaga gcaaaccaga gtgcttggag tatggtaact tggaacagag
     2461 gtgactccat tcagacacag caagagaccc acttagaaac acgcctttat tatctgcacg
     2521 gagcccagac ttgtcctcag tccttgggag agggagtctt tgagagtggt gggaggtctt
     2581 gggtcatagt ctccacatac catgagggca ggaaggcata gggtgcgtca cgatggacag
     2641 acacagtgac agggccacca ggttcccagg agaagaggtg gacaagcctg tggaagacag
     2701 aggtgtgggg tgaagccaga gattgagccg ggatccagag gcagtgcaca aacatgaagg
     2761 ccttacctgg gatcgtcctg gactactgtg ctctgggcaa agctaaggaa ggcagcacag
     2821 aagttcatgc acacagaggg aattcagcct tctcggtcat tctggagggg gcacaagagg
     2881 cagaagactg ggggtggaaa gaggaaaagg caggtggcaa aggggcactg ggtctgagga
     2941 cttggggccg gccggatttt tgtaaaagag agaatgttta aaagttttgt tactttatag
     3001 aagaaatttt gagtttttgt ttttttttaa taaataaata aacataaata aattgtttgt
     3061 tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta ttcaaattaa
     3121 taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg catgattatc
     3181 tttaacgtac gtcacaatat gattatcttt ctagggttaa ataatagttt ctaatttttt
     3241 tattattcag gcgaagttac atgtatcgat ctcgaggggg ggcccggtac ccaattcgcc
     3301 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     3361 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     3421 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3481 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3541 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3601 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3661 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3721 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3781 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3841 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3901 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     3961 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4021 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4081 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4141 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4201 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4261 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4321 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4381 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4441 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4501 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4561 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4621 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4681 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4741 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4801 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4861 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4921 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4981 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5041 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5101 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5161 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5221 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5281 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5341 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5401 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5461 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5521 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5581 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5641 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5701 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5761 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5821 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     5881 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     5941 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6001 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     6061 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcga aattaaccct
     6121 cactaaaggg aacaaaagct ggagctccac cgcggaaagt ttaggtcgag taaagcgcaa
     6181 atctttttta accctagaaa gatagtctgc gtaaaattga cgcatgcatt cttgaaatat
     6241 tgctctctct ttctaaatag cgcgaatccg tcgctgtgca tttaggacat ctcagtcgcc
     6301 gcttggagct cccgtgaggc gtgcttgtca atgcggtaag tgtcactgat tttgaactat
     6361 aacgaccgcg tgagtcaaaa tgacgcatga ttatctttta cgtgactttt aagatttaac
     6421 tcatacgata attatattgt tatttcatgt tctacttacg tgataactta ttatatatat
     6481 attttcttgt tatagatgcc taccggccga gccaaggaga cagactgggc agccccctga
     6541 gtgggtattt ctacagggtg acgacttagg ttgaaaccca tatggaactt ctagacacaa
     6601 aactaaagac catccaccac cttctacaag actgccagcc tggcacagga gtggaagttc
     6661 cagaaccagg taaaaggttg ggaggtttct atggctggag agccatggct gtgaaactag
     6721 agcaagttgg tctctccttc acagaggcca gcataccctt tactgaggac ctggggaaag
     6781 aagaccaaga agctggaaac agtcaggcct gggaagagcc cataacctgg agcccagaaa
     6841 cacggaacct agctccttcc tgggaggtgg agcaggggct gaggagaagg tggcacaaga
     6901 cggtgacaaa gggtccagca gtaaatgggg aacaaccact gaaggtttag taaaaagagt
     6961 cccctcgtac tgggtgttcc tctgtgcttt ttccactccc tgctacacac acacacacgg
     7021 tgcaaaagcc aagcactctt cccttattag gaagagagag actgccttcc agaatgtgaa
     7081 tggagaagac agagagagag ccacagttct gcccttgtgt ttctgaggct accgagatgc
     7141 cagccatcaa ggctcaaact ccattttctc gtgatttaga caggaaggct tttgcaaaaa
     7201 caacccttta ttaagatcca gtggggaaat aaaggcgcga gcttgatatc gaattaattc
     7261 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     7321 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     7381 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     7441 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     7501 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     7561 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     7621 aggggtgggt ccggggg
//
