LOCUS       sequence-267610-        6852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human SOX17 upon doxycycline treatment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  AUTHORS   Ream MW, Randolph LN, Jiang Y, Chang Y, Bao X, Lian XL
  TITLE     Direct programming of human pluripotent stem cells into endothelial 
            progenitors with SOX17 and FGF2.
  JOURNAL   Stem Cell Reports. 2024 Mar 5:S2213-6711(24)00044-4. doi: 
            10.1016/j.stemcr.2024.02.006.
  PUBMED    38518781
REFERENCE   2  (bases 1 to 6852)
  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: "Stem Cell 
            Reports. 2024 Mar 5:S2213-6711(24)00044-4. doi: 
            10.1016/j.stemcr.2024.02.006."
COMMENT     Sequence Label: XLone-SOX17
FEATURES             Location/Qualifiers
     source          1..6852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(361..384)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      384..393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(413..624)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        666..1033
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    674..692
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    710..728
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    746..764
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    782..800
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    818..836
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    854..872
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    890..908
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1994..2011
                     /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             2000..2017
                     /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"
     polyA_signal    2471..2605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2881..2915
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2926..2943)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2926..2942)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2935..2957)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3151..3170)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3270..3292
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3330..3348)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3416..3520
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3521..3589,3590..4381)
                     /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(3739..3758)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4552..5140
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5041..5060
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5294..5311
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5428..5449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5464..5469,5470..5487,5488..5494)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5502..5518
                     /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     5507..5529
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5526..5542
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5526..5542
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5568..5630
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    5806..5887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(6004..6402)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(6403..6459)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(6469..6852,1..360))
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
ORIGIN
        1 caggaacgcg agctgatttt ccagggtttc gtactgtttc tctgttgggc gggtgccgag
       61 atgcacttta gccccgtcgc gatgtgagag gagagcacag cggtatgact tggcgttgtt
      121 ccgcagaaag tcttgccatg actcgccttc cagggggcag gagtgggtat gatgcctgtc
      181 cagcatctcg attggcaggg catcgagcag ggcccgcttg ttcttcacgt gccagtacag
      241 ggtaggctgc tcaactccca gcttttgagc gagtttcctt gtcgtcaggc cttcgatacc
      301 gactccattg agtaattcca gagcagagtt tatgactttg ctcttgtcca gtctagacat
      361 cttatcgtca tcgtctttgt aatccatggt ggcggatccc gcgtcacgac acctgtgttc
      421 tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt atatacggtt
      481 ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc cagtttaccc
      541 cgcgccacct tctctaggca ccggttcaat tgccgacccc tccccccaac ttctcgggga
      601 ctgtgggcga tgtgcgctct gcccactgac gggcaccgga gccactcgag tggaattatc
      661 acctcgagtt tactccctat cagtgataga gaacgtatga agagtttact ccctatcagt
      721 gatagagaac gtatgcagac tttactccct atcagtgata gagaacgtat aaggagttta
      781 ctccctatca gtgatagaga acgtatgacc agtttactcc ctatcagtga tagagaacgt
      841 atctacagtt tactccctat cagtgataga gaacgtatat ccagtttact ccctatcagt
      901 gatagagaac gtataagctt tgcttatgta aaccagggcg cctataaaag agtgctgatt
      961 ttttgagtaa acttcaattc cacaacactt ttgtcttata ccaactttcc gtaccacttc
     1021 ctaccctcgt aaaggtaccg ccaccatgag cagcccggat gcgggatacg ccagtgacga
     1081 ccagagccag acccagagcg cgctgcccgc ggtgatggcc gggctgggcc cctgcccctg
     1141 ggccgagtcg ctgagcccca tcggggacat gaaggtgaag ggcgaggcgc cggcgaacag
     1201 cggagcaccg gccggggccg cgggccgagc caagggcgag tcccgtatcc ggcggccgat
     1261 gaacgctttc atggtgtggg ctaaggacga gcgcaagcgg ctggcgcagc agaatccaga
     1321 cctgcacaac gccgagttga gcaagatgct gggcaagtcg tggaaggcgc tgacgctggc
     1381 ggagaagcgg cccttcgtgg aggaggcaga gcggctgcgc gtgcagcaca tgcaggacca
     1441 ccccaactac aagtaccggc cgcggcggcg caagcaggtg aagcggctga agcgggtgga
     1501 gggcggcttc ctgcacggcc tggctgagcc gcaggcggcc gcgctgggcc ccgagggcgg
     1561 ccgcgtggcc atggacggcc tgggcctcca gttccccgag cagggcttcc ccgccggccc
     1621 gccgctgctg cctccgcaca tgggcggcca ctaccgcgac tgccagagtc tgggcgcgcc
     1681 tccgctcgac ggctacccgt tgcccacgcc cgacacgtcc ccgctggacg gcgtggaccc
     1741 cgacccggct ttcttcgccg ccccgatgcc cggggactgc ccggcggccg gcacctacag
     1801 ctacgcgcag gtctcggact acgctggccc cccggagcct cccgccggtc ccatgcaccc
     1861 ccgactcggc ccagagcccg cgggtccctc gattccgggc ctcctggcgc cacccagcgc
     1921 ccttcacgtg tactacggcg cgatgggctc gcccggggcg ggcggcgggc gcggcttcca
     1981 gatgcagccg caacaccagc accagcacca gcaccagcac caccccccgg gccccggaca
     2041 gccgtcgccc cctccggagg cactgccctg ccgggacggc acggacccca gtcagcccgc
     2101 cgagctcctc ggggaggtgg accgcacgga atttgaacag tatctgcact tcgtgtgcaa
     2161 gcctgagatg ggcctcccct accaggggca tgactccggt gtgaatctcc ccgacagcca
     2221 cggggccatt tcctcggtgg tgtccgacgc cagctccgcg gtatattact gcaactatcc
     2281 tgacgtgtag actagtagac cacctcccct gcgagctaag ctggacagcc aatgacgggt
     2341 aagagagtga catttttcac taacctaaga caggagggcc gtcagagcta ctgcctaatc
     2401 caaagacggg taaaagtgat aaaaatgtat cactccaacc taagacaggc gcagcttccg
     2461 agggatttga gatccagaca tgataagata cattgatgag tttggacaaa ccaaaactag
     2521 aatgcagtga aaaaaatgcc ttatttgtga aatttgtgat gctattgcct tatttgtaac
     2581 cattataagc tgcaataaac aagtttgata tctataacaa gaaaatatat atataataag
     2641 ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa
     2701 gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca
     2761 gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt
     2821 cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg
     2881 catgcgtcaa ttttacgcag actatctttc tagggttaag aattcactgg ccgtcgtttt
     2941 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3001 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3061 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3121 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3181 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3241 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3301 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3361 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3421 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3481 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3541 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3601 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3661 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3721 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3781 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3841 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3901 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3961 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4021 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4081 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4141 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4201 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4261 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4321 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4381 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4441 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4501 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4561 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4621 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4681 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4741 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4801 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4861 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4921 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4981 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5041 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5101 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5161 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5221 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5281 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5341 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5401 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     5461 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5521 tcacacagga aacagctatg accatgatta cgccaaggtc gacttaaccc tagaaagata
     5581 atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg tgttttatcg
     5641 gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca
     5701 cttacatact aataataaat tcaacaaaca atttatttat gtttatttat ttattaaaaa
     5761 aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttagcagt gaaaaaaatg
     5821 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     5881 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     5941 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctgga
     6001 gatttagccc tcccacacat aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg
     6061 ctgtccatca ctgtccttca ctatggcttt gatcccagga tgcagatcga gaagcacctg
     6121 tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca tttccgatcg cgacgataca
     6181 agtcaggttg ccagctgccg cagcagcagc agtgcccagc accacgagtt ctgcacaagg
     6241 tcccccagta aaatgatata cattgacacc agtgaagatg cggccgtcgc tagagagagc
     6301 tgcgctggcg acgctgtagt cttcagagat ggggatgctg ttgattgtag ccgttgctct
     6361 ttcaatgagg gtggattctt cttgagacaa aggcttggcc atggggccgg ggttctcctc
     6421 cacgtcgccg gcctgcttca gcaggctgaa gttggtggcg ccgctgcccc cggggagcat
     6481 gtcaaggtca aaatcgtcaa gagcgtcagc aggcagcata tcaaggtcaa agtcgtcaag
     6541 ggcatcggct gggagcatgt ctaagtcaaa atcgtcaagg gcgtcggtcg gcccgccgct
     6601 ttcgcacttt agctgtttct ccaggccaca tatgattagt tccaggccga aaaggaaggc
     6661 aggttcggct ccctgccggt cgaacagctc aattgcttgt ttcagaagtg ggggcataga
     6721 atcggtggta ggtgtctctc tttcctcttt tgctacttga tgctcctgtt cctccaatac
     6781 gcagcccagt gtaaagtggc ccacggcgga cagagcgtac agtgcgttct ccagggagaa
     6841 gccttgctga ca
//
