LOCUS       sequence-465408-        7529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Salpingoeca helianthica full-length Sox-II in mammalian 
            cells, for lentivirus generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7529)
  AUTHORS   Gao Y, Tan DS, Girbig M, Hu H, Zhou X, Xie Q, Yeung SW, Lee KS, Ho 
            SY, Cojocaru V, Yan J, Hochberg GKA, de Mendoza A, Jauch R
  TITLE     The emergence of Sox and POU transcription factors predates the 
            origins of animal stem cells.
  JOURNAL   Nat Commun. 2024 Nov 14;15(1):9868. doi: 10.1038/s41467-024-54152-x.
  PUBMED    39543096
REFERENCE   2  (bases 1 to 7529)
  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: "Nat 
            Commun."; date: "2024-11-14"; volume: "15(1):9868. doi"; pages: " 
            10.1038/s41467-024-54152-x"
COMMENT     Sequence Label: pHAGE2-TetO-Salhel-Sox-II
FEATURES             Location/Qualifiers
     source          1..7529
                     /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    3937..4525
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3990..4010)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4408..4419)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4655..4688)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             4707..4887
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4934..4952)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5020..5124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5125..5193,5194..5985)
                     /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(5343..5362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6156..6744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6898..6915
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7155..7176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7191..7196,7197..7214,7215..7221)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7229..7245
                     /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     7234..7256
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7253..7269
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7253..7269
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7319..7338
                     /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 atgccaggaa
     2701 gaggaccaag agctagagaa agtgataaag caagactcgc cttctctcac aaggcaggag
     2761 aagacaatca ggccctggcc acaatgaaca tgactatcgg cggcttcttc ctgctgagtg
     2821 agccagacga cctgggtggc cacggccacc acagtaacga cacaatgggt gacctgcccg
     2881 agcatctgtt cggcgctcct aatagtctcc tgagcgattc tcccgatgcc tctagcacca
     2941 tgggtgcagt ggcagccgct gctgccgccg cagcagccgc agctgctgcc gccgcaagga
     3001 accaaaatcg ccctggtcag ctccctccaa gatctgctaa ccatgtgaag cgccctatga
     3061 atgctttcat ggtttgggct aaagaacaga ggcgcgagta ccggaggaca caccctaacg
     3121 ctcacaatgc tgccgtgagc aaatatctcg gacagaggtg gaatgagctg agtcctagtg
     3181 tgaaagagga atacgcagag cgggctagaa tggtcaaaat cgagcatgcc gcagccaatc
     3241 caggctacaa atactgccca cggcgcatgc tccctggcag accaggcagc cctacacgcg
     3301 caacaacacg cggactgctg gacgctgccg ggcctgatat gccacagttc gccatgcctg
     3361 gagccgggcc aagtctgaga atgccttccg gtctgctcct gtccctggga gatggagctg
     3421 gcgctccact gcagctgtat gatggaagca ttgcagctgg aggacaggag gaggccgggc
     3481 tcagacatga cggcggtggt tctgtgcccg atggttccct ggctgatccc ttcggaaatg
     3541 cttgtagcta tcctgttctg gctcctacac agcctagcct cgatcaacct gcagccgatc
     3601 acatgccagc tccattcgct cctatgccac tgctgcctcc actgcctcct ctcccagccc
     3661 tcgacaggga gccctcttcc gctgcaagaa tggccgacgc cctgctggca acaagtctgg
     3721 gactccctga gctgtctgtg ggcgatgttg aagacgtcct cggtgtgtcc gcttgtgccc
     3781 tgcccgcaga cttcacactg gatctcacac tgaatcacca cctctccatt acatcttgtc
     3841 agggacagct ggtgctcgac gacttttctc aaagcactac cttttctctc gctcggaacc
     3901 agaatggctc cgcataagtc gacatcgata gatcctaatc aacctctgga ttacaaaatt
     3961 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4021 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     4081 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     4141 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4201 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     4261 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     4321 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     4381 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     4441 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     4501 atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc
     4561 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     4621 acgaagacaa gatcacctgc aggacaggcg cgccataact tcgtatagca tacattatac
     4681 gaagttatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     4741 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     4801 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     4861 tttagtcagt gtggaaaatc tctagcaccc gggcgattaa ggaaagggct agatcattct
     4921 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4981 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5041 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5101 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5161 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5221 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5281 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5341 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     5401 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5461 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5521 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5581 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5641 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5701 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5761 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5821 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5881 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5941 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6001 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6061 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6301 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6781 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6841 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6901 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     6961 tggccgattc attaatgcag caagctcatg gctgactaat tttttttatt tatgcagagg
     7021 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7081 taggcttttg caaaaagctc cccgtggcac gacaggtttc ccgactggaa agcgggcagt
     7141 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     7201 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7261 agctatgaca tgattacgaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7321 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct
     7381 aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg
     7441 tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt
     7501 tgttaaatat gtactacaaa cttagtagt
//
