LOCUS       sequence-410022-        7979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP fused mouse SOX2 in mammalian cells, for lentivirus 
            generation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7979)
  AUTHORS   Hu H, Ho DHH, Tan DS, MacCarthy CM, Yu CH, Weng M, Scholer HR, Jauch
            R
  TITLE     Evaluation of the determinants for improved pluripotency induction 
            and maintenance by engineered SOX17.
  JOURNAL   Nucleic Acids Res. 2023 Aug 23:gkad597. doi: 10.1093/nar/gkad597.
  PUBMED    37607832
REFERENCE   2  (bases 1 to 7979)
  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: "Nucleic 
            Acids Res. 2023 Aug 23:gkad597. doi: 10.1093/nar/gkad597."
COMMENT     Sequence Label: pHAGE2-TetO-GFP-mSox2
FEATURES             Location/Qualifiers
     source          1..7979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2251..2269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2293..2311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2335..2353
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2377..2395
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2419..2437
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2461..2479
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2547..2571
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2649..2651,2652..2654,2655..3365)
                     /codon_start=1
                     /product="enhanced GFP (EGFP) derivative that exhibits 
                     robust folding and enhanced photostability (Zhong et al., 
                     2019)"
                     /label=SiriusGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2694..2715)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3302..3323
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4387..4975
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4440..4460)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4858..4869)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5105..5138)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5157..5337
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5384..5402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5470..5574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5575..5643,5644..6435)
                     /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(5793..5812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6606..7194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7348..7365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7605..7626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7641..7646,7647..7664,7665..7671)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7679..7695
                     /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     7684..7706
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7703..7719
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7703..7719
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7769..7788
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcca
     2221 caccctaact gacacactcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     2281 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     2341 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     2401 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     2461 tccctatcag tgatagagaa aagtgaaagt cgagctcggt acccgggtcg aggtaggcat
     2521 gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
     2581 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctgcggcca
     2641 ccaccggtat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     2701 agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc gagggcgatg
     2761 ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2821 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     2881 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2941 ccatctcctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg
     3001 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     3061 tggggcacaa gctggagtac aacttcaaca gccacaacgt ctatatcacg gccgacaagc
     3121 agaagaacgg catcaaggcg aacttcaaga tccgccacaa cgtcgaggac ggcagcgtgc
     3181 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3241 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     3301 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3361 acaagggtgc accgggttct gcgggttctg ccgcaggtgg atccggtatg tataacatga
     3421 tggagacgga gctgaagccg ccgggcccgc agcaagcttc ggggggcggc ggcggaggag
     3481 gcaacgccac ggcggcggcg accggcggca accagaagaa cagcccggac cgcgtcaaga
     3541 ggcccatgaa cgccttcatg gtatggtccc gggggcagcg gcgtaagatg gcccaggaga
     3601 accccaagat gcacaactcg gagatcagca agcgcctggg cgcggagtgg aaacttttgt
     3661 ccgagaccga gaagcggccg ttcatcgacg aggccaagcg gctgcgcgct ctgcacatga
     3721 aggagcaccc ggattataaa taccggccgc ggcggaaaac caagacgctc atgaagaagg
     3781 ataagtacac gcttcccgga ggcttgctgg cccccggcgg gaacagcatg gcgagcgggg
     3841 ttggggtggg cgccggcctg ggtgcgggcg tgaaccagcg catggacagc tacgcgcaca
     3901 tgaacggctg gagcaacggc agctacagca tgatgcagga gcagctgggc tacccgcagc
     3961 acccgggcct caacgctcac ggcgcggcac agatgcaacc gatgcaccgc tacgacgtca
     4021 gcgccctgca gtacaactcc atgaccagct cgcagaccta catgaacggc tcgcccacct
     4081 acagcatgtc ctactcgcag cagggcaccc ccggtatggc gctgggctcc atgggctctg
     4141 tggtcaagtc cgaggccagc tccagccccc ccgtggttac ctcttcctcc cactccaggg
     4201 cgccctgcca ggccggggac ctccgggaca tgatcagcat gtacctcccc ggcgccgagg
     4261 tgccggagcc cgctgcgccc agtagactgc acatggccca gcactaccag agcggcccgg
     4321 tgcccggcac ggccattaac ggcacactgc ccctgtcgca catgtgagtc gacatcgata
     4381 gatcctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4441 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4501 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4561 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4621 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4681 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4741 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     4801 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4861 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4921 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt
     4981 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     5041 ggggggactg gaagggctaa ttcactccca acgaagacaa gatcacctgc aggacaggcg
     5101 cgccataact tcgtatagca tacattatac gaagttatct gctttttgct tgtactgggt
     5161 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5221 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5281 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaccc
     5341 gggcgattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt gatacgccta
     5401 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5461 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5521 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5581 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5641 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5701 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5761 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5821 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5881 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5941 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6001 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6061 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6121 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6181 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6241 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6301 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6361 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6421 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6481 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6541 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6601 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6661 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6721 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6781 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6841 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6901 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6961 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7021 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7081 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7141 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7201 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7261 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7321 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7381 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag caagctcatg
     7441 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     7501 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc cccgtggcac
     7561 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7621 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7681 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa tttcacaaat
     7741 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     7801 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
     7861 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
     7921 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
