LOCUS       sequence-410020-        8321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP fused mouse SOX17 in mammalian cells, for lentivirus 
            generation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8321)
  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 8321)
  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-mSox17
FEATURES             Location/Qualifiers
     source          1..8321
                     /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    join(2251..2269,2270..2292,2293..2311,2312..2334,2335..2353,
                     2354..2376,2377..2395,2396..2418,2419..2437,2438..2460,
                     2461..2479,2480..2502,2503..2521)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     primer_bind     2589..2613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2691..2693,2694..2696,2697..3407)
                     /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(2736..2757)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3344..3365
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4729..5317
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4782..4802)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5200..5211)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5447..5480)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5499..5679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5726..5744)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5812..5916
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5917..5985,5986..6777)
                     /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(6135..6154)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6948..7536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7690..7707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7947..7968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7983..7988,7989..8006,8007..8013)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8021..8037
                     /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     8026..8048
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8045..8061
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8045..8061
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8111..8130
                     /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 cgagtttacc actccctatc agtgatagag
     2521 aaaagtgaaa gtcgagctcg gtacccgggt cgaggtaggc atgtacggtg ggaggcctat
     2581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     2641 gacctccata gaagacaccg ggaccgatcc agcctgcggc caccaccggt atggtgagca
     2701 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2761 acggccacaa gttcagcgtg cgcggcgagg gcgagggcga tgccaccaac ggcaagctga
     2821 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2881 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2941 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctcc ttcaaggacg
     3001 acggcaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     3061 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3121 acaacttcaa cagccacaac gtctatatca cggccgacaa gcagaagaac ggcatcaagg
     3181 cgaacttcaa gatccgccac aacgtcgagg acggcagcgt gcagctcgcc gaccactacc
     3241 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     3301 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3361 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggt gcaccgggtt
     3421 ctgcgggttc tgccgcaggt ggatccggta tgagcagccc ggatgcggga tacgccagtg
     3481 acgaccagag ccagccccgg agcgcgcagc ccgcggtgat ggcagggttg ggcccctgtc
     3541 cctgggccga gtccctgagc cccctcgggg atgtaaaggt gaaaggcgag gtggtggcga
     3601 gtagcggggc gccagccggg acgtcgggcc gagccaaagc ggagtctcgc atccggcggc
     3661 cgatgaacgc ctttatggtg tgggccaaag acgaacgcaa gcggttggca cagcagaacc
     3721 cagatctgca caacgcagag ctaagcaaga tgctaggcaa gtcttggaag gcgttgacct
     3781 tggcagagaa gcggcccttc gtggaagagg ccgagcggct gcgcgtgcag catatgcagg
     3841 accaccccaa ctacaagtac cggccgcggc ggcgcaagca ggtgaagcgc atgaagcggg
     3901 tggagggagg cttcctgcac gctctcgtcg agccccaggc cggcgcgctt ggtcccgagg
     3961 gcggccgcgt ggccatggat ggcctgggtc tgcctttccc ggagccgggc tatccggccg
     4021 gtcctccgct gatgtctccg cacatgggcc cccactatcg ggactgccag ggactgggcg
     4081 ctcccgcgct cgacggctac cctctgccca ctccggacac atccccgctg gatggcgtgg
     4141 agcaggaccc ggctttcttt gcagccccgc tgccagggga ctgcccggcg gccggcacct
     4201 acacttacgc tccagtctcg gactatgcag tgtccgtaga gccgcccgct ggccccatgc
     4261 gagtggggcc ggacccctcg ggccctgcga tgccggggat cctggcgccc cccagcgctc
     4321 tgcacctgta ctacggcgcg atgggctcgc ccgccgcaag tgcggggcgc ggtttccacg
     4381 cgcaacccca gcagccgctg caaccgcagg caccgccccc gccaccgcag cagcagcacc
     4441 cagcgcacgg ccccgggcaa ccttcgcccc ctcccgaggc tctgccctgc cgggatggca
     4501 cggaatccaa ccagcccact gagctcctag gggaggtgga ccgcacggaa ttcgaacagt
     4561 atctgccctt tgtgtataag cccgagatgg gtcttcccta ccagggacac gactgcggag
     4621 tgaacctctc agacagccac ggagccattt cctccgtggt gtccgacgct agctcagcgg
     4681 tctactattg caactacccc gacatttaag tcgacatcga tagatcctaa tcaacctctg
     4741 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4801 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4861 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4921 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4981 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5041 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5101 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     5161 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5221 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5281 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac
     5341 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5401 aattcactcc caacgaagac aagatcacct gcaggacagg cgcgccataa cttcgtatag
     5461 catacattat acgaagttat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     5521 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     5581 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     5641 ccctcagacc cttttagtca gtgtggaaaa tctctagcac ccgggcgatt aaggaaaggg
     5701 ctagatcatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5761 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5821 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5881 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5941 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6001 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6061 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6121 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     6181 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6241 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6301 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6361 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6421 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6481 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6541 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6601 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6661 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6721 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6781 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6841 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6901 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7081 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7501 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7561 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7621 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7681 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7741 ctccccgcgc gttggccgat tcattaatgc agcaagctca tggctgacta atttttttta
     7801 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     7861 tttttggagg cctaggcttt tgcaaaaagc tccccgtggc acgacaggtt tcccgactgg
     7921 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7981 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     8041 cacacaggaa acagctatga catgattacg aatttcacaa ataaagcatt tttttcactg
     8101 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg
     8161 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc
     8221 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
     8281 agaaattgta tttgttaaat atgtactaca aacttagtag t
//
