LOCUS       sequence-97518-e        5442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac expression vector encoding murine Sox17 downstream of a 
            doxycycline inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5442)
  AUTHORS   Vereide DT, Vickerman V, Swanson SA, Chu LF, McIntosh BE, Thomson JA
  TITLE     An expandable, inducible hemangioblast state regulated by fibroblast
            growth factor.
  JOURNAL   Stem Cell Reports. 2014 Dec 9;3(6):1043-57. doi: 
            10.1016/j.stemcr.2014.10.003. Epub 2014 Nov 13.
  PUBMED    25458896
REFERENCE   2  (bases 1 to 5442)
  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"; doi: 
            "10.1016/j.stemcr.2014.10.003"; journalName: "Stem Cell Reports"; 
            date: "2014-12-9- 9"; volume: "3"; issue: "6"; pages: "1043-57"
COMMENT     Sequence Label: PB-Sox17
FEATURES             Location/Qualifiers
     source          1..5442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..81
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    453..471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    490..508
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    527..545
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    564..582
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    601..619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    638..656
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    675..693
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    712..730
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     752..776
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    856..880
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2174..2198)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2271..2288)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2277..2388
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2762..2796
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2822..2838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2822..2838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2835..2857)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2846..2862
                     /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(2870..2876,2877..2894,2895..2900))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2915..2936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3053..3070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3224..3812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3304..3323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3983..4774,4775..4843))
                     /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     4606..4625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4844..4948)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5016..5034
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5072..5094)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5194..5213
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5407..5429
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5421..5438
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5422..5438
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcaagcttta ttatttaacc ctagaaagat aatcatattg tgacgtacgt taaagataat
       61 catgcgtaaa attgacgcat gtgttttatc ggtctgtata tcgaggttta tttattaatt
      121 tgaatagata ttaagtttta ttatatttac acttacatac taataataaa ttcaacaaac
      181 aatttattta tgtttattta tttattaaaa aaaaacaaaa actcaaaatt tcttctataa
      241 agtaacaaaa cttttaaaca ttctctcttt tacaaaaata aacttatttt gtactttaaa
      301 aacagtcatg ttgtaatcga taataagtaa ttagcttaac ctatacataa tagaaacaaa
      361 ttatacttat tagtcagtca gaaacaactt tggcacatat caatattatg ctctcgttaa
      421 ttaactcgag ggatccaagc tggccgcacg tctccctatc agtgatagag aagtcgacac
      481 gtctcgagct ccctatcagt gatagagaag gtacgtctag aacgtctccc tatcagtgat
      541 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtacgt ctagaacgtc
      601 tccctatcag tgatagagaa gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt
      661 acgtctagaa cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca
      721 gtgatagaga aggtaccccc tatataagca gagctcgttt agtgaaccgt cagatcgcct
      781 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
      841 gcggtggcgg ccatcacaag tttgtacaaa aaagcaggct tgaaggaatt cggtaccatg
      901 agcagcccgg atgcgggata cgccagtgac gaccagagcc agccccggag cgcgcagccc
      961 gcggtgatgg cagggttggg cccctgtccc tgggccgagt ccctgagccc cctcggggat
     1021 gtaaaggtga aaggcgaggt ggtggcgagt agcggggcgc cagccgggac gtcgggccga
     1081 gccaaagcgg agtctcgcat ccggcggccg atgaacgcct ttatggtgtg ggccaaagac
     1141 gaacgcaagc ggttggcaca gcagaaccca gatctgcaca acgcagagct aagcaagatg
     1201 ctaggcaagt cttggaaggc gttgaccttg gcagagaagc ggcccttcgt ggaagaggcc
     1261 gagcggctgc gcgtgcagca tatgcaggac caccccaact acaagtaccg gccgcggcgg
     1321 cgcaagcagg tgaagcgcat gaagcgggtg gagggaggct tcctgcacgc tctcgtcgag
     1381 ccccaggccg gcgcgcttgg tcccgagggc ggccgcgtgg ccatggatgg cctgggtctg
     1441 cctttcccgg agccgggcta tccggccggt cctccgctga tgtctccgca catgggcccc
     1501 cactatcggg actgccaggg actgggcgct cccgcgctcg acggctaccc tctgcccact
     1561 ccggacacat ccccgctgga tggcgtggag caggacccgg ctttctttgc agccccgctg
     1621 ccaggggact gcccggcggc cggcacctac acttacgctc cagtctcgga ctatgcagtg
     1681 tccgtagagc cgcccgctgg ccccatgcga gtggggccgg acccctcggg ccctgcgatg
     1741 ccggggatcc tggcgccccc cagcgctctg cacctgtact acggcgcgat gggctcgccc
     1801 gccgcaagtg cggggcgcgg tttccacgcg caaccccagc agccgctgca accgcaggca
     1861 ccgcccccgc caccgcagca gcagcaccca gcgcacggcc ccgggcaacc ttcgccccct
     1921 cccgaggctc tgccctgccg ggatggcacg gaatccaacc agcccactga gctcctaggg
     1981 gaggtggacc gcacggaatt cgaacagtat ctgccctttg tgtataagcc cgagatgggt
     2041 cttccctacc agggacacga ctgcggagtg aacctctcag acagccacgg agccatttcc
     2101 tccgtggtgt ccgacgctag ctcagcggtc tactattgca actaccccga catttagctc
     2161 gagtgcggcc gcaacccagc tttcttgtac aaagtggtga tccaaccggt atagatctat
     2221 ggccggccat caattgatac tagtcaccga ctctagagct cgctgatcag cctcgactgt
     2281 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2341 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2401 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2461 agacaatagc aggcactagt cggtcacgat atctataaca agaaaatata tatataataa
     2521 gttatcacgt aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa
     2581 agtcacgtaa aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc
     2641 agtgacactt accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg
     2701 tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat
     2761 gcatgcgtca attttacgca gactatcttt ctagggttaa aaaaagcttg gcgtaatcat
     2821 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2881 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2941 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3001 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3061 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3121 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3181 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3241 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3301 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3361 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3421 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3481 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3541 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3601 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3661 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3721 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3781 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3841 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3901 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3961 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4021 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4081 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4141 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4201 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4261 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4321 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4381 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4441 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4501 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4561 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4621 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4681 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4741 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4801 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4861 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4921 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4981 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5041 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5101 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5161 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     5221 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     5281 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     5341 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     5401 ggttttccca gtcacgacgt tgtaaaacga cggccagtga at
//
