LOCUS       sequence-410037-        7256 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses human SOX2 in mammalian cells, for lentivirus generation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7256)
  AUTHORS   Weng M, Hu H, Graus MS, Tan DS, Gao Y, Ren S, Ho DHH, Langer J, 
            Holzner M, Huang Y, Ling GS, Lai CSW, Francois M, Jauch R
  TITLE     An engineered Sox17 induces somatic to neural stem cell fate 
            transitions independently from pluripotency reprogramming.
  JOURNAL   Sci Adv. 2023 Aug 25;9(34):eadh2501. doi: 10.1126/sciadv.adh2501. 
            Epub 2023 Aug 23.
  PUBMED    37611093
REFERENCE   2  (bases 1 to 7256)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2023-08-25"; volume: "9(34):eadh2501. doi"; pages: " 
            10.1126/sciadv.adh2501. Epub 2023 Aug 23"
COMMENT     Sequence Label: pHAGE2-TetO-hSOX2
FEATURES             Location/Qualifiers
     source          1..7256
                     /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"
     misc_feature    3664..4252
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3717..3737)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4135..4146)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4382..4415)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             4434..4614
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4661..4679)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4747..4851
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4852..4920,4921..5712)
                     /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(5070..5089)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5883..6471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6625..6642
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6882..6903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6918..6923,6924..6941,6942..6948)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6956..6972
                     /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     6961..6983
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6980..6996
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6980..6996
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7046..7065
                     /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 atgtacaaca
     2701 tgatggagac ggagctgaag ccgccgggcc cgcagcaaac ttcggggggc ggcggcggca
     2761 actccaccgc ggcggcggcc ggcggcaacc agaaaaacag cccggaccgc gtcaagcggc
     2821 ccatgaatgc cttcatggtg tggtcccgcg ggcagcggcg caagatggcc caagagaacc
     2881 ccaagatgca caactcggag atcagcaagc gcctgggcgc cgagtggaaa cttttgtcgg
     2941 agacggagaa gcggccgttc atcgacgagg ctaagcggct gcgagcgctg cacatgaagg
     3001 agcacccgga ttataaatac cggccccggc ggaaaaccaa gacgctcatg aagaaggata
     3061 agtacacgct gcccggcggg ctgctggccc ccggcggcaa tagcatggcg agcggggtcg
     3121 gggtgggcgc cggcctgggc gcgggcgtga accagcgcat ggacagttac gcgcacatga
     3181 acggctggag caacggcagc tacagcatga tgcaggacca gctgggctac ccgcagcacc
     3241 cgggcctcaa tgcgcacggc gcagcgcaga tgcagcccat gcaccgctac gacgtgagcg
     3301 ccctgcagta caactccatg accagctcgc agacctacat gaacggctcg cccacctaca
     3361 gcatgtccta ctcgcagcag ggcacccctg gcatggctct tggctccatg ggttcggtgg
     3421 tcaagtccga ggccagctcc agcccccctg tggttacctc ttcctcccac tccagggcgc
     3481 cctgccaggc cggggacctc cgggacatga tcagcatgta tctccccggc gccgaggtgc
     3541 cggaacccgc cgcccccagc agacttcaca tgtcccagca ctaccagagc ggcccggtgc
     3601 ccggcacggc cattaacggc acactgcccc tctcacacat gtgagtcgac atcgatagat
     3661 cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3721 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3781 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3841 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3901 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3961 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4021 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     4081 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4141 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4201 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc
     4261 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     4321 gggactggaa gggctaattc actcccaacg aagacaagat cacctgcagg acaggcgcgc
     4381 cataacttcg tatagcatac attatacgaa gttatctgct ttttgcttgt actgggtctc
     4441 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     4501 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     4561 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcacccggg
     4621 cgattaagga aagggctaga tcattcttga agacgaaagg gcctcgtgat acgcctattt
     4681 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4741 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4801 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4861 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4921 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4981 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5041 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     5101 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5161 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5221 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5281 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5341 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5401 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5461 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5521 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5581 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5641 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5701 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5761 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5821 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5881 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5941 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6001 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6061 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6121 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6181 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6241 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6301 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6361 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6421 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6481 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6541 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6601 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6661 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa gctcatggct
     6721 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6781 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc gtggcacgac
     6841 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6901 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6961 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt cacaaataaa
     7021 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7081 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
     7141 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
     7201 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt
//
