LOCUS       sequence-320848-        6762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human SOX18 upon doxycycline treatment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6762)
  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 6762)
  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-SOX18
FEATURES             Location/Qualifiers
     source          1..6762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    37..55
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    73..91
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    109..127
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    145..163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    181..199
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    217..235
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     polyA_signal    1708..1842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2118..2152
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2163..2180)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2163..2179)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2172..2194)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2388..2407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2507..2529
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2567..2585)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2653..2757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2758..2826,2827..3618)
                     /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(2976..2995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3789..4377
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4278..4297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4531..4548
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4665..4686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4701..4706,4707..4724,4725..4731)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4739..4755
                     /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     4744..4766
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4763..4779
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4763..4779
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   4805..4867
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    5043..5124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5241..5639)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(5640..5696)
                     /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(5706..6449)
                     /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"
     CDS             complement(6450..6473)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      6473..6482
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6502..6713)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        join(6755..6762,1..360)
                     /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"
ORIGIN
        1 tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta
       61 tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg
      121 atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac
      181 tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta
      241 taagctttgc ttatgtaaac cagggcgcct ataaaagagt gctgattttt tgagtaaact
      301 tcaattccac aacacttttg tcttatacca actttccgta ccacttccta ccctcgtaaa
      361 ggtaccgcca ccatgcagag atcgccgccc ggctacggcg cacaggacga cccgcccgcc
      421 cgccgcgact gtgcatgggc cccgggacac ggggccgccg ctgacacgcg cggcctcgcc
      481 gccggccccg ccgccctcgc cgcgcccgcc gcgcccgcct cgccgcccag cccgcagcgc
      541 agtcccccgc gcagccccga gccggggcgc tatggcctca gcccggccgg ccgcggggaa
      601 cgccaggcgg cagacgagtc gcgcatccgg cggcccatga acgccttcat ggtgtgggca
      661 aaggacgagc gcaagcggct ggctcagcag aacccggacc tgcacaacgc ggtgctcagc
      721 aagatgctgg gcaaagcgtg gaaggagctg aacgcggcgg agaagcggcc cttcgtggag
      781 gaagccgaac ggctgcgcgt gcagcacttg cgcgaccacc ccaactacaa gtaccggccg
      841 cgccgcaaga agcaggcgcg caaggcccgg cggctggagc ccggcctcct gctcccggga
      901 ttagcgcccc cgcagccacc gcccgagcct ttccccgcgg cgtctggctc ggctcgcgcc
      961 ttccgcgagc tgcccccgct gggcgccgag ttcgacggcc tggggctgcc cacgcccgag
     1021 cgctcgcctc tggacggcct ggagcccggc gaggctgcct tcttcccacc gcccgcggcg
     1081 cccgaggact gcgcgctgcg gcccttccgc gcgccctacg cgcccaccga gttgtcgcgg
     1141 gaccccggcg gttgctacgg ggctcccctg gcggaggcgc tcaggaccgc gccccccgcg
     1201 gcgccgctcg ctggcctgta ctacggcacc ctgggcacgc ccggcccgta ccccggcccg
     1261 ctgtcgccgc cgcccgaggc cccgccgctg gagagcgccg agccgctggg gcccgccgcc
     1321 gatctgtggg ccgacgtgga cctcaccgag ttcgaccagt acctcaactg cagccggact
     1381 cggcccgacg cccccgggct cccgtaccac gtggcactgg ccaaactggg cccgcgcgcc
     1441 atgtcctgcc cagaggagag cagcctgatc tccgcgctgt cggacgccag cagcgcggtc
     1501 tattacagcg cgtgcatctc cggctagact agtagaccac ctcccctgcg agctaagctg
     1561 gacagccaat gacgggtaag agagtgacat ttttcactaa cctaagacag gagggccgtc
     1621 agagctactg cctaatccaa agacgggtaa aagtgataaa aatgtatcac tccaacctaa
     1681 gacaggcgca gcttccgagg gatttgagat ccagacatga taagatacat tgatgagttt
     1741 ggacaaacca aaactagaat gcagtgaaaa aaatgcctta tttgtgaaat ttgtgatgct
     1801 attgccttat ttgtaaccat tataagctgc aataaacaag tttgatatct ataacaagaa
     1861 aatatatata taataagtta tcacgtaagt agaacatgaa ataacaatat aattatcgta
     1921 tgagttaaat cttaaaagtc acgtaaaaga taatcatgcg tcattttgac tcacgcggtc
     1981 gttatagttc aaaatcagtg acacttaccg cattgacaag cacgcctcac gggagctcca
     2041 agcggcgact gagatgtcct aaatgcacag cgacggattc gcgctattta gaaagagaga
     2101 gcaatatttc aagaatgcat gcgtcaattt tacgcagact atctttctag ggttaagaat
     2161 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2221 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2281 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     2341 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     2401 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2461 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2521 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     2581 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2641 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2701 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2761 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2821 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2881 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2941 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3001 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3061 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3121 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3181 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3241 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3301 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3361 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3421 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3481 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3541 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3601 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3661 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3721 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3781 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3841 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3901 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3961 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4021 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4081 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4141 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4201 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4261 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4321 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4381 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4441 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4501 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4561 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4621 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     4681 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     4741 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caaggtcgac
     4801 ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg cgtaaaattg
     4861 acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa tagatattaa
     4921 gttttattat atttacactt acatactaat aataaattca acaaacaatt tatttatgtt
     4981 tatttattta ttaaaaaaaa acaaaaactc aaaatttctt ctataaagta acaaaacttt
     5041 tagcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     5101 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     5161 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     5221 gattatgatc ctctggagat ttagccctcc cacacataac cagagggcag caattcacga
     5281 atcccaactg ccgtcggctg tccatcactg tccttcacta tggctttgat cccaggatgc
     5341 agatcgagaa gcacctgtcg gcaccgtccg caggggctca agatgcccct gttctcattt
     5401 ccgatcgcga cgatacaagt caggttgcca gctgccgcag cagcagcagt gcccagcacc
     5461 acgagttctg cacaaggtcc cccagtaaaa tgatatacat tgacaccagt gaagatgcgg
     5521 ccgtcgctag agagagctgc gctggcgacg ctgtagtctt cagagatggg gatgctgttg
     5581 attgtagccg ttgctctttc aatgagggtg gattcttctt gagacaaagg cttggccatg
     5641 gggccggggt tctcctccac gtcgccggcc tgcttcagca ggctgaagtt ggtggcgccg
     5701 ctgcccccgg ggagcatgtc aaggtcaaaa tcgtcaagag cgtcagcagg cagcatatca
     5761 aggtcaaagt cgtcaagggc atcggctggg agcatgtcta agtcaaaatc gtcaagggcg
     5821 tcggtcggcc cgccgctttc gcactttagc tgtttctcca ggccacatat gattagttcc
     5881 aggccgaaaa ggaaggcagg ttcggctccc tgccggtcga acagctcaat tgcttgtttc
     5941 agaagtgggg gcatagaatc ggtggtaggt gtctctcttt cctcttttgc tacttgatgc
     6001 tcctgttcct ccaatacgca gcccagtgta aagtggccca cggcggacag agcgtacagt
     6061 gcgttctcca gggagaagcc ttgctgacac aggaacgcga gctgattttc cagggtttcg
     6121 tactgtttct ctgttgggcg ggtgccgaga tgcactttag ccccgtcgcg atgtgagagg
     6181 agagcacagc ggtatgactt ggcgttgttc cgcagaaagt cttgccatga ctcgccttcc
     6241 agggggcagg agtgggtatg atgcctgtcc agcatctcga ttggcagggc atcgagcagg
     6301 gcccgcttgt tcttcacgtg ccagtacagg gtaggctgct caactcccag cttttgagcg
     6361 agtttccttg tcgtcaggcc ttcgataccg actccattga gtaattccag agcagagttt
     6421 atgactttgc tcttgtccag tctagacatc ttatcgtcat cgtctttgta atccatggtg
     6481 gcggatcccg cgtcacgaca cctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt
     6541 cacggcgact actgcactta tatacggttc tcccccaccc tcgggaaaaa ggcggagcca
     6601 gtacacgaca tcactttccc agtttacccc gcgccacctt ctctaggcac cggttcaatt
     6661 gccgacccct ccccccaact tctcggggac tgtgggcgat gtgcgctctg cccactgacg
     6721 ggcaccggag ccactcgagt ggaattatca cctcgagttt ac
//
