LOCUS       sequence-200843-        6425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector encoding doxycycline-inducible human SOX17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6425)
  AUTHORS   Miller AZ, Satchie A, Tannenbaum AP, Nihal A, Thomson JA, Vereide DT
  TITLE     Expandable Arterial Endothelial Precursors from Human CD34(+) Cells 
            Differ in Their Proclivity to Undergo an Endothelial-to-Mesenchymal 
            Transition.
  JOURNAL   Stem Cell Reports. 2018 Jan 9;10(1):73-86. doi: 
            10.1016/j.stemcr.2017.12.011.
  PUBMED    29320761
REFERENCE   2  (bases 1 to 6425)
  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.2017.12"; journalName: "Stem Cell Reports"; date: 
            "2018-01-9- 9"; volume: "10"; issue: "1"; pages: "73-86"
COMMENT     Sequence Label: PB-TRE3G-SOX17
FEATURES             Location/Qualifiers
     source          1..6425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..53)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     153..172
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     366..388
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     380..397
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     381..397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   420..482
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1430..1808
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1438..1456
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1474..1492
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1510..1528
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1546..1564
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1582..1600
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1618..1636
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1654..1672
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1717..1741
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1826..1850
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2816..2833
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     CDS             2822..2839
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     protein_bind    complement(3129..3153)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3203..3324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3214..3233)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3268..3287
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(3795..3828)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     repeat_region   4146..4180
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4206..4222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4206..4222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4219..4241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4230..4246
                     /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(4254..4260,4261..4278,4279..4284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4299..4320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4437..4454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4608..5196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4688..4707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5367..6158,6159..6227))
                     /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     5990..6009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6228..6332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6400..6418
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
       61 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
      121 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
      181 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
      241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      361 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcaagcttt attatttaac
      421 cctagaaaga taatcatatt gtgacgtacg ttaaagataa tcatgcgtaa aattgacgca
      481 tgtgttttat cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt
      541 attatattta cacttacata ctaataataa attcaacaaa caatttattt atgtttattt
      601 atttattaaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa acttttaaac
      661 attctctctt ttacaaaaat aaacttattt tgtactttaa aaacagtcat gttgtaatcg
      721 ataataagta attagcttaa cctatacata atagaaacaa attatactta ttagtcagtc
      781 agaaacaact ttggcacata tcaatattat gctctcgtta attaactcga gggatccggc
      841 gcgcctgtca ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt
      901 gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc
      961 cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc
     1021 ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc
     1081 cctgggggct ttgggagggg gctgtccccg tgagctccca ggcgcgcctg tcattctaaa
     1141 tctctctttc agcctaaagc tttttccccg tatcccccca ggtgtctgca ggctcaaaga
     1201 gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc
     1261 cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg agccccgggc
     1321 ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg gctttggggg
     1381 ggggctgtcc ccgtgagctc ccaggcgcgc cgctagccaa gaattcctcg agtttactcc
     1441 ctatcagtga tagagaacgt atgaagagtt tactccctat cagtgataga gaacgtatgc
     1501 agactttact ccctatcagt gatagagaac gtataaggag tttactccct atcagtgata
     1561 gagaacgtat gaccagttta ctccctatca gtgatagaga acgtatctac agtttactcc
     1621 ctatcagtga tagagaacgt atatccagtt tactccctat cagtgataga gaacgtataa
     1681 gctttaggcg tgtacggtgg gcgcctataa aagcagagct cgtttagtga accgtcagat
     1741 cgcctggagc aattccacaa cacttttgtc ttataccaac tttccgtacc acttcctacc
     1801 ctcgtaaacc gcggtggcgg ccatcacaag tttgtacaaa aaagcaggct tggaaggagt
     1861 tcgaaccatg agcagcccgg atgcgggata cgccagtgac gaccagagcc agacccagag
     1921 cgcgctgccc gcggtgatgg ccgggctggg cccctgcccc tgggccgagt cgctgagccc
     1981 catcggggac atgaaggtga agggcgaggc gccggcgaac agcggagcac cggccggggc
     2041 cgcgggccga gccaagggcg agtcccgtat ccggcggccg atgaacgctt tcatggtgtg
     2101 ggctaaggac gagcgcaagc ggctggcgca gcagaatcca gacctgcaca acgccgagtt
     2161 gagcaagatg ctgggcaagt cgtggaaggc gctgacgctg gcggagaagc ggcccttcgt
     2221 ggaggaggca gagcggctgc gcgtgcagca catgcaggac caccccaact acaagtaccg
     2281 gccgcggcgg cgcaagcagg tgaagcggct gaagcgggtg gagggcggct tcctgcacgg
     2341 cctggctgag ccgcaggcgg ccgcgctggg ccccgagggc ggccgcgtgg ccatggacgg
     2401 cctgggcctc cagttccccg agcagggctt ccccgccggc ccgccgctgc tgcctccgca
     2461 catgggcggc cactaccgcg actgccagag tctgggcgcg cctccgctcg acggctaccc
     2521 gttgcccacg cccgacacgt ccccgctgga cggcgtggac cccgacccgg ctttcttcgc
     2581 cgccccgatg cccggggact gcccggcggc cggcacctac agctacgcgc aggtctcgga
     2641 ctacgctggc cccccggagc ctcccgccgg tcccatgcac ccccgactcg gcccagagcc
     2701 cgcgggtccc tcgattccgg gcctcctggc gccacccagc gcccttcacg tgtactacgg
     2761 cgcgatgggc tcgcccgggg cgggcggcgg gcgcggcttc cagatgcagc cgcaacacca
     2821 gcaccagcac cagcaccagc accacccccc gggccccgga cagccgtcgc cccctccgga
     2881 ggcactgccc tgccgggacg gcacggaccc cagtcagccc gccgagctcc tcggggaggt
     2941 ggaccgcacg gaatttgaac agtatctgca cttcgtgtgc aagcctgaga tgggcctccc
     3001 ctaccagggg catgactccg gtgtgaatct ccccgacagc cacggggcca tttcctcggt
     3061 ggtgtccgac gccagctccg cggtatatta ctgcaactat cctgacgtgt agctcgagtg
     3121 cggccgcaac ccagctttct tgtacaaagt ggtgatccaa ccggtataga tctcgagctc
     3181 gatgagttta aaccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3241 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3301 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3361 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     3421 gattatgatc cggctgcctc gcgcttaatt aacccggggg ccgcctggga actgccgctg
     3481 gccccccacc gccccaagga tctccggtcc ccgcccggct tgctgacgtc acggatgccc
     3541 cagggtgtgc tgggcaggta gcgggagcct gggaaatgga gtccattagc agaagtggcc
     3601 acttccagga gtcgctgtgc cccgatgcac actgggaagt ccgcagctcc gaggccccca
     3661 gtggaaatcg ccagatgagg gcctcctccg gggaatgctg ggaaatggag tctacaggcc
     3721 ggaggggtcc ccacggcata ctagcaagtg tgtgtagcac cgcgtaaagg ggatgaatag
     3781 cagactgccc cgggataact tcgtatagga tactttatac gaagttatga gctctgtgat
     3841 aatcactagt gaattgatac tagtcggtca cgatatctat aacaagaaaa tatatatata
     3901 ataagttatc acgtaagtag aacatgaaat aacaatataa ttatcgtatg agttaaatct
     3961 taaaagtcac gtaaaagata atcatgcgtc attttgactc acgcggtcgt tatagttcaa
     4021 aatcagtgac acttaccgca ttgacaagca cgcctcacgg gagctccaag cggcgactga
     4081 gatgtcctaa atgcacagcg acggattcgc gctatttaga aagagagagc aatatttcaa
     4141 gaatgcatgc gtcaatttta cgcagactat ctttctaggg ttaaaaaaag cttggcgtaa
     4201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6361 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6421 tttcg
//
