LOCUS       sequence-431754-        7648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generate lentiviruses encoding for human SOX17E57C (HMG box 
            numbering); for use in pluripotency reprogramming.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7648)
  AUTHORS   Ho SY, Hu H, Ho DHH, Renom APS, Yeung SW, Boerner F, Weng M, 
            Hutchins AP, Jauch R
  TITLE     An acidic residue within the OCT4 dimerization interface of SOX17 is
            necessary and sufficient to overcome its pluripotency-inducing 
            activity.
  JOURNAL   Stem Cell Reports. 2025 Mar 11;20(3):102398. doi: 
            10.1016/j.stemcr.2025.102398. Epub 2025 Feb 6.
  PUBMED    39919754
REFERENCE   2  (bases 1 to 7648)
  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."; date: "2025-03-11"; volume: "20(3):102398. doi"; pages: "
            10.1016/j.stemcr.2025.102398. Epub 2025 Feb 6"
COMMENT     Sequence Label: pHAGE2-TetO-SOX17E57C
FEATURES             Location/Qualifiers
     source          1..7648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(174..192)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        260..364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(365..433,434..1225)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(583..602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1396..1984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2138..2155
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2395..2416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2431..2436,2437..2454,2455..2461)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2469..2485
                     /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     2474..2496
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2493..2509
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2493..2509
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2559..2578
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             2770..3403
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3450..3575
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4072..4305
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4490..4534
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4827..4944
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    5020..5038
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5062..5080
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5104..5122
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5146..5164
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     gap             5176..5275
                     /estimated_length=100
     primer_bind     5332..5356
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             6382..6399
                     /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             6388..6405
                     /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"
     misc_feature    6698..7286
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6751..6771)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7169..7180)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(7416..7449)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             7468..7648
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
       61 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
      121 tctagcaccc gggcgattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt
      181 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      241 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      301 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      361 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      421 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      481 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      541 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      601 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      661 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      721 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      781 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      841 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      901 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      961 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1021 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1081 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1141 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1201 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1261 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1321 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1381 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1441 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1501 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1561 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1621 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1681 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1741 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1801 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1861 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1921 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1981 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2041 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2101 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2161 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2221 caagctcatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2281 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     2341 cccgtggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2401 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     2461 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
     2521 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2581 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact
     2641 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg
     2701 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa
     2761 cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga
     2821 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca
     2881 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag
     2941 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg
     3001 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg
     3061 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg
     3121 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag
     3181 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac
     3241 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     3301 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     3361 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     3421 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     3481 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     3541 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     3601 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     3661 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     3721 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     3781 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     3841 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     3901 ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     3961 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     4021 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     4081 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     4141 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     4201 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     4261 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     4321 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     4381 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     4441 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     4501 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     4561 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     4621 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     4681 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     4741 agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccgaat
     4801 tcacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     4861 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     4921 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     4981 actagtccac accctaactg acacactcga gtttaccact ccctatcagt gatagagaaa
     5041 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
     5101 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
     5161 gagaaaagtg aaagtnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     5221 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnncgagc
     5281 tcggtacccg ggtcgaggta ggcatgtacg gtgggaggcc tatataagca gagctcgttt
     5341 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
     5401 ccgggaccga tccagcctgc ggccaccacc ggtatgagca gcccggatgc gggatacgcc
     5461 agtgacgacc agagccagac ccagagcgcg ctgcccgcgg tgatggccgg gctgggcccc
     5521 tgcccctggg ccgagtcgct gagccccatc ggggacatga aggtgaaggg cgaggcgccg
     5581 gcgaacagcg gagcaccggc cggggccgcg ggccgagcca agggcgagtc ccgtatccgg
     5641 cggccgatga acgctttcat ggtgtgggct aaggacgagc gcaagcggct ggcgcagcag
     5701 aatccagacc tgcacaacgc cgagttgagc aagatgctgg gcaagtcgtg gaaggcgctg
     5761 acgctggcgg aaaagcggcc cttcgtggag gaagcatgcc ggctgcgcgt gcagcacatg
     5821 caggaccacc ccaactacaa gtaccggccg cggcggcgca agcaggtgaa gcggctgaag
     5881 cgggtggagg gcggcttcct gcacggcctg gctgagccgc aggcggccgc gctgggcccc
     5941 gagggcggcc gcgtggccat ggacggcctg ggcctccagt tccccgagca gggcttcccc
     6001 gccggcccgc cgctgctgcc tccgcacatg ggcggccact accgcgactg ccagagtctg
     6061 ggcgcgcctc cgctcgacgg ctacccgttg cccacgcccg acacgtcccc gctggacggc
     6121 gtggaccccg acccggcttt cttcgccgcc ccgatgcccg gggactgccc ggcggccggc
     6181 acctacagct acgcgcaggt ctcggactac gctggccccc cggagcctcc cgccggtccc
     6241 atgcaccccc gactcggccc agagcccgcg ggtccctcga ttccgggcct cctggcgcca
     6301 cccagcgccc ttcacgtgta ctacggcgcg atgggctcgc ccggggcggg cggcgggcgc
     6361 ggcttccaga tgcagccgca acaccagcac cagcaccagc accagcacca ccccccgggc
     6421 cccggacagc cgtcgccccc tccggaggca ctgccctgcc gggacggcac ggaccccagt
     6481 cagcccgccg agctcctcgg ggaggtggac cgcacggaat ttgaacagta tctgcacttc
     6541 gtgtgcaagc ctgagatggg cctcccctac caggggcatg actccggtgt gaatctcccc
     6601 gacagccacg gggccatttc ctcggtggtg tccgacgcca gctccgcggt atattactgc
     6661 aactatcctg acgtgtgagt cgacatcgat agatcctaat caacctctgg attacaaaat
     6721 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6781 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     6841 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     6901 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     6961 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     7021 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     7081 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     7141 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     7201 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     7261 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     7321 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     7381 aacgaagaca agatcacctg caggacaggc gcgccataac ttcgtatagc atacattata
     7441 cgaagttatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     7501 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7561 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7621 ttttagtcag tgtggaaaat ctctagca
//
