LOCUS       sequence-320850-        6774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human SOX7 upon doxycycline treatment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6774)
  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 6774)
  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-SOX7
FEATURES             Location/Qualifiers
     source          1..6774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..368
                     /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    9..27
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    45..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    81..99
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    117..135
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    153..171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    189..207
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    225..243
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      375..384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1728..1862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2138..2172
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2183..2200)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2183..2199)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2192..2214)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2408..2427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2527..2549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2587..2605)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2673..2777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2778..2846,2847..3638)
                     /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(2996..3015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3809..4397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4298..4317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4551..4568
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4685..4706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4721..4726,4727..4744,4745..4751)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4759..4775
                     /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     4764..4786
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4783..4799
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4783..4799
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   4825..4887
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    5063..5144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5261..5659)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(5660..5716)
                     /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(5726..6469)
                     /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(6470..6493)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      6493..6502
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6522..6733)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag
       61 agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc
      121 tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta
      181 cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag
      241 agaacgtata agctttgctt atgtaaacca gggcgcctat aaaagagtgc tgattttttg
      301 agtaaacttc aattccacaa cacttttgtc ttataccaac tttccgtacc acttcctacc
      361 ctcgtaaagg taccgccacc atggcttcgc tgctgggagc ctacccttgg cccgagggtc
      421 tcgagtgccc ggccctggac gccgagctgt cggatggaca atcgccgccg gccgtccccc
      481 ggcccccggg ggacaagggc tccgagagcc gtatccggcg gcccatgaac gccttcatgg
      541 tttgggccaa ggacgagagg aaacggctgg cagtgcagaa cccggacctg cacaacgccg
      601 agctcagcaa gatgctggga aagtcgtgga aggcgctgac gctgtcccag aagaggccgt
      661 acgtggacga ggcggagcgg ctgcgcctgc agcacatgca ggactacccc aactacaagt
      721 accggccgcg caggaagaag caggccaagc ggctgtgcaa gcgcgtggac ccgggcttcc
      781 ttctgagctc cctctcccgg gaccagaacg ccctgccgga gaagagaagc ggcagccggg
      841 gggcgctggg ggagaaggag gacaggggtg agtactcccc cggcactgcc ctgcccagcc
      901 tccggggctg ctaccacgag gggccggctg gtggtggcgg cggcggcacc ccgagcagtg
      961 tggacacgta cccgtacggg ctgcccacac ctcctgaaat gtctcccctg gacgtgctgg
     1021 agccggagca gaccttcttc tcctccccct gccaggagga gcatggccat ccccgccgca
     1081 tcccccacct gccagggcac ccgtactcac cggagtacgc cccaagccct ctccactgta
     1141 gccaccccct gggctccctg gcccttggcc agtcccccgg cgtctccatg atgtcccctg
     1201 tacccggctg tcccccatct cctgcctatt actccccggc cacctaccac ccactccact
     1261 ccaacctcca agcccacctg ggccagcttt ccccgcctcc tgagcaccct ggcttcgacg
     1321 ccctggatca actgagccag gtggaactcc tgggggacat ggatcgcaat gaattcgacc
     1381 agtatttgaa cactcctggc cacccagact ccgccacagg ggccatggcc ctcagtgggc
     1441 atgttccggt ctcccaggtg acaccaacgg gtcccacaga gaccagcctc atctccgtcc
     1501 tggctgatgc cacggccacg tactacaaca gctacagtgt gtcatagact agtagaccac
     1561 ctcccctgcg agctaagctg gacagccaat gacgggtaag agagtgacat ttttcactaa
     1621 cctaagacag gagggccgtc agagctactg cctaatccaa agacgggtaa aagtgataaa
     1681 aatgtatcac tccaacctaa gacaggcgca gcttccgagg gatttgagat ccagacatga
     1741 taagatacat tgatgagttt ggacaaacca aaactagaat gcagtgaaaa aaatgcctta
     1801 tttgtgaaat ttgtgatgct attgccttat ttgtaaccat tataagctgc aataaacaag
     1861 tttgatatct ataacaagaa aatatatata taataagtta tcacgtaagt agaacatgaa
     1921 ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga taatcatgcg
     1981 tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg cattgacaag
     2041 cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag cgacggattc
     2101 gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt tacgcagact
     2161 atctttctag ggttaagaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2221 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2281 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     2341 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     2401 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     2461 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2521 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     2581 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2641 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2701 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2761 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2821 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2881 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2941 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3001 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3061 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3121 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3181 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3241 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3301 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3361 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3421 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3481 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3541 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3601 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3661 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3721 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3781 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3901 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3961 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4021 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4081 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4141 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4201 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4261 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4321 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4381 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4441 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4501 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4561 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4621 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4681 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     4741 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     4801 atgattacgc caaggtcgac ttaaccctag aaagataatc atattgtgac gtacgttaaa
     4861 gataatcatg cgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta
     4921 ttaatttgaa tagatattaa gttttattat atttacactt acatactaat aataaattca
     4981 acaaacaatt tatttatgtt tatttattta ttaaaaaaaa acaaaaactc aaaatttctt
     5041 ctataaagta acaaaacttt tagcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     5101 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     5161 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     5221 tctacaaatg tggtatggct gattatgatc ctctggagat ttagccctcc cacacataac
     5281 cagagggcag caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta
     5341 tggctttgat cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca
     5401 agatgcccct gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag
     5461 cagcagcagt gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat
     5521 tgacaccagt gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt
     5581 cagagatggg gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt
     5641 gagacaaagg cttggccatg gggccggggt tctcctccac gtcgccggcc tgcttcagca
     5701 ggctgaagtt ggtggcgccg ctgcccccgg ggagcatgtc aaggtcaaaa tcgtcaagag
     5761 cgtcagcagg cagcatatca aggtcaaagt cgtcaagggc atcggctggg agcatgtcta
     5821 agtcaaaatc gtcaagggcg tcggtcggcc cgccgctttc gcactttagc tgtttctcca
     5881 ggccacatat gattagttcc aggccgaaaa ggaaggcagg ttcggctccc tgccggtcga
     5941 acagctcaat tgcttgtttc agaagtgggg gcatagaatc ggtggtaggt gtctctcttt
     6001 cctcttttgc tacttgatgc tcctgttcct ccaatacgca gcccagtgta aagtggccca
     6061 cggcggacag agcgtacagt gcgttctcca gggagaagcc ttgctgacac aggaacgcga
     6121 gctgattttc cagggtttcg tactgtttct ctgttgggcg ggtgccgaga tgcactttag
     6181 ccccgtcgcg atgtgagagg agagcacagc ggtatgactt ggcgttgttc cgcagaaagt
     6241 cttgccatga ctcgccttcc agggggcagg agtgggtatg atgcctgtcc agcatctcga
     6301 ttggcagggc atcgagcagg gcccgcttgt tcttcacgtg ccagtacagg gtaggctgct
     6361 caactcccag cttttgagcg agtttccttg tcgtcaggcc ttcgataccg actccattga
     6421 gtaattccag agcagagttt atgactttgc tcttgtccag tctagacatc ttatcgtcat
     6481 cgtctttgta atccatggtg gcggatcccg cgtcacgaca cctgtgttct ggcggcaaac
     6541 ccgttgcgaa aaagaacgtt cacggcgact actgcactta tatacggttc tcccccaccc
     6601 tcgggaaaaa ggcggagcca gtacacgaca tcactttccc agtttacccc gcgccacctt
     6661 ctctaggcac cggttcaatt gccgacccct ccccccaact tctcggggac tgtgggcgat
     6721 gtgcgctctg cccactgacg ggcaccggag ccactcgagt ggaattatca cctc
//
