LOCUS       sequence-465592-        8057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Salpingoeca helianthica full-length Sox-I in mammalian 
            cells, for lentivirus generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8057)
  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 8057)
  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-I
FEATURES             Location/Qualifiers
     source          1..8057
                     /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             3774..3791
                     /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    4465..5053
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4518..4538)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4936..4947)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5183..5216)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5235..5415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5462..5480)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5548..5652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5653..5721,5722..6513)
                     /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(5871..5890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6684..7272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7426..7443
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7683..7704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7719..7724,7725..7742,7743..7749)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7757..7773
                     /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     7762..7784
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7781..7797
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7781..7797
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7847..7866
                     /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 atggataagg
     2701 aaccagcact cggctttgtc gaatctgcca ataaagcata tcctctgttt actctggatg
     2761 aaatggcagc tagcgtgctg gatacagcat ccgctagaga aagcctggcc gcctccgagc
     2821 tgcctgccaa atctccagcc agtttctccc tgaaaagagg ccacgaagag ctcagtcagg
     2881 acgatgcctc tccctccagg gtgcccaccc attccatggt cgcaccagca ccaggtgctg
     2941 caggctctgg aggcggcagt gagccaccat ttgtgcgcag gcagcgggct aatagcctgt
     3001 gctttggtaa ttttacctct tccaacagag agccaatcgc caggccactg gtgtccatcg
     3061 tgctgggtag tcctcctcca ccagaaaagg cctcccacat caaacggccc atgaacgcct
     3121 tcatggtgtg ggctcaggag agaagaaggg aagttaaaat gtgtaatcct accctccaca
     3181 atgctaccat cagtagaagc ctcggagagg aatggaatgc cctgcctgag gcagagaagc
     3241 agccatttgt ggagagagcc agggctctga aggccgagca tcaacaagaa cacccagatt
     3301 acaaatacaa tccacggaga aaacctaagc tggacaacgc tcatactctc agccctctgg
     3361 tgacttcctc cgtgtgtatg cctgctgcaa tcaagacact cacaccacct cggagaaggg
     3421 ccgccagctt ttctactcct tctgctgcta gatgtgagag aggagtcgtg gcagggtcca
     3481 atctggccca gactacaagc ctgtgtgacg ccatgctcga tctgctcgga cctacattcg
     3541 ccaacctgac ctctgctaac gcacataata tgcctggtct ggtcccatcc tctcatctgc
     3601 atgaccatgc taatcgcgat ggtgatacat ccgtgctgcc cgcttttgat cccgctgttt
     3661 tccctctcga tgggttcatg cctgaaactc ggtccgccct ctacgtcatg gacaatacta
     3721 ctacagggac aaccaaccac catcatcaca acgcctactg ccagcaccaa caccatcaac
     3781 accagcacca gcatcatcat cagcaccatg ccggctccgg agtgtctttc tcttgcgggc
     3841 acatatatca tctgagcaca agtccacgcg ccagcggagc tggcatgtct acaagcgggt
     3901 ctagcggcga cctcgcatct gcagcctctc tcagggtttg tggagccaat gccacaggag
     3961 aggaaaatcc ctgctgcggc ggtctgggtt gtggcggacg cggatgtgat gctgccatct
     4021 ttgcacgggt tcctcctgca actccttcta gaaccctgca cacccggccc acaccaccac
     4081 gctcccaccc agttcacacc tctcatgccc acgtccggtc taacgcatcc gccggcaatc
     4141 attcctttac actggagagc cctagcaggt gcctggaaag cctgctcatg tctcaagctc
     4201 ctgaaaccgc accaataacc aacactgcca cacagtctga cgcaaccgca gctcacaccc
     4261 ataatcacgc aagagccgga gaggcagatg caacctggga gacagtggag ctgtgtggag
     4321 ctggcgttac aagtggtatt gccatactca ccggcagcgc ccagcccaac gctccactgg
     4381 aggtcgatgt gggtatggtg gattggggta caattgaaca gctcctcgat gtgtggcctc
     4441 tgtaagtcga catcgataga tcctaatcaa cctctggatt acaaaatttg tgaaagattg
     4501 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4561 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4621 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4681 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4741 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4801 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4861 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4921 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4981 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     5041 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     5101 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     5161 tcacctgcag gacaggcgcg ccataacttc gtatagcata cattatacga agttatctgc
     5221 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5281 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5341 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5401 ggaaaatctc tagcacccgg gcgattaagg aaagggctag atcattcttg aagacgaaag
     5461 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5521 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5581 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5641 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5701 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5761 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5821 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5881 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5941 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6001 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6061 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6121 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6181 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6241 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6301 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6361 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6421 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6481 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6541 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6601 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6661 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6721 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6781 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6841 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6901 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6961 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7021 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7081 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7141 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7201 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7261 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7321 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7381 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7441 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7501 taatgcagca agctcatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7561 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     7621 aaaagctccc cgtggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7681 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7741 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgacatg
     7801 attacgaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     7861 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
     7921 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
     7981 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
     8041 actacaaact tagtagt
//
