LOCUS       sequence-465442-        7223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Mylnosiga fluctuans full-length POU in mammalian cells, 
            for lentivirus generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7223)
  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 7223)
  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-Myflu-POU
FEATURES             Location/Qualifiers
     source          1..7223
                     /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    3631..4219
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3684..3704)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4102..4113)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4349..4382)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             4401..4581
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4628..4646)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4714..4818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4819..4887,4888..5679)
                     /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(5037..5056)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5850..6438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6592..6609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6849..6870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6885..6890,6891..6908,6909..6915)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6923..6939
                     /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     6928..6950
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6947..6963
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6947..6963
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7013..7032
                     /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 atgagcggca
     2701 tggagagcaa ccctgccatg gagggcccta cccacgccga catcgtggac gagggcgccg
     2761 agatcggccc taccatgagc aaggagatgc aagaggctag gaggctggac agcattagga
     2821 ccatcttcca catgaccggc ctgatgctgc tgagggccgc cgaccctaac cctgccgccg
     2881 acctgagcgt gagcatcgct cagctgagcc aaaccgccga gaccatcatc cctaacctgc
     2941 ctaacatcct gaggacctac ggcctggaga gcctggagcc tctgatcacc gaccctgaga
     3001 tcctggtgaa cgtgacccaa gacatcctgg cctgcgtgac cgccggccct cctcaagaga
     3061 acagcaacct gatcgccctg aggaagaagc tggccgccct ggcccctagg gccgacgaca
     3121 gcgacgacgt gggcatggag aggctgagga agttcgccac cttcctgcaa gagtggagga
     3181 gcagccacct gagccaaatg cagttcctgg acctgatgca cgcccactgc cctgtgaagt
     3241 acacacagac ctgcctgagc agattcgaga agggcaacct gagctatcag agcatgagga
     3301 ggatgcacgg cttcatcgag aagaccatgc tggcctacgc cgagaaccct ttcgccgccc
     3361 ctgccaccgt gaggaagagg aagcagaggg agaccatcag catccaagtg aggcagcagc
     3421 tggaggtgca cttccaagct agccctaggc ctagcctgga cgccctgcaa gtgatcgcta
     3481 gggacctggg cctcagcctg gagacaacaa ggtactggtt tcagaacgct aggagcagca
     3541 acaagaagca gaggcctgac agcgtggaga tcggctgtag gcctgagcct ggccctgagg
     3601 acgctagcta agtcgacatc gatagatcct aatcaacctc tggattacaa aatttgtgaa
     3661 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3721 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3781 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3841 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3901 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3961 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4021 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4081 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4141 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4201 ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     4261 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     4321 acaagatcac ctgcaggaca ggcgcgccat aacttcgtat agcatacatt atacgaagtt
     4381 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     4441 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     4501 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     4561 cagtgtggaa aatctctagc acccgggcga ttaaggaaag ggctagatca ttcttgaaga
     4621 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4681 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4741 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4801 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4861 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4921 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4981 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5041 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     5101 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5161 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5221 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5281 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5341 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5401 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5461 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5521 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5581 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5641 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5701 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5761 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5821 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5881 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5941 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6001 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6061 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6481 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6541 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6601 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6661 attcattaat gcagcaagct catggctgac taattttttt tatttatgca gaggccgagg
     6721 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6781 tttgcaaaaa gctccccgtg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6841 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6901 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     6961 gacatgatta cgaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     7021 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
     7081 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
     7141 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
     7201 atatgtacta caaacttagt agt
//
