LOCUS       sequence-172690-        8682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8682)
  AUTHORS   Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, 
            Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II, 
            Thomson JA
  TITLE     Induced pluripotent stem cell lines derived from human somatic 
            cells.
  JOURNAL   Science. 2007 Dec 21. 318(5858):1917-20.
  PUBMED    18029452
REFERENCE   2  (bases 1 to 8682)
  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: "Science. 
            2007 Dec 21. 318(5858):1917-20."
COMMENT     Sequence Label: pSin-EF2-Oct4-Pur
FEATURES             Location/Qualifiers
     source          1..8682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             117..750
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    797..922
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1415..1648
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1833..1877
                     /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    2164..2281
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2409..3587
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2640..3578
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3535..3555
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    join(4798..5372,5373..5375,5376..5384)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4965..4982)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5192..5211
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5404..6003
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(5404..5423)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5900..5920
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6206..6439
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6590..6607)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6596..6820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      6863..6872
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        6916..7020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7021..7089,7090..7881)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7239..7258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8052..8640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8541..8560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggaccgcca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
       61 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttgga
      121 agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct accacacaca
      181 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
      241 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
      301 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
      361 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
      421 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
      481 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
      541 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
      601 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      661 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      721 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      781 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      841 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      901 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
      961 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1021 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1081 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1141 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1201 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1261 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1321 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1381 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1441 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1501 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1561 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1621 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1681 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     1741 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     1801 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     1861 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     1921 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     1981 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     2041 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     2101 gatccattcg attagtgaac ggatctcgac ggtatcgcca caaatggcag tattcatcca
     2161 caattttaaa agaaaggggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     2221 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     2281 tcgggtttat tacagggaca gcagagatcc actttggtcg ataagctttg caaagatgga
     2341 taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag
     2401 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     2461 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
     2521 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     2581 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     2641 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
     2701 cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
     2761 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
     2821 aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
     2881 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
     2941 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
     3001 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
     3061 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
     3121 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
     3181 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
     3241 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
     3301 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     3361 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     3421 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     3481 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     3541 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgagga attgatcctt
     3601 cgaactagtg attatggcgg gacacctggc ttcggatttc gccttctcgc cccctccagg
     3661 tggtggaggt gatgggccag gggggccgga gccgggctgg gttgatcctc ggacctggct
     3721 aagcttccaa ggccctcctg gagggccagg aatcgggccg ggggttgggc caggctctga
     3781 ggtgtggggg attcccccat gccccccgcc gtatgagttc tgtgggggga tggcgtactg
     3841 tgggccccag gttggagtgg ggctagtgcc ccaaggcggc ttggagacct ctcagcctga
     3901 gggtgaagca ggagtcgggg tggagagcaa ctccgatggg gcctccccgg agccctgcac
     3961 cgtcacccct ggtgccgtga agctggagaa ggagaagctg gagcaaaacc cggaggagtc
     4021 ccaggacatc aaagctctgc agaaagaact cgagcaattt gccaagctcc tgaagcagaa
     4081 gaggatcacc ctgggatata cacaggccga tgtggggctc accctggggg ttctatttgg
     4141 gaaggtattc agccaaacga ccatctgccg ctttgaggct ctgcagctta gcttcaagaa
     4201 catgtgtaag ctgcggccct tgctgcagaa gtgggtggag gaagctgaca acaatgaaaa
     4261 tcttcaggag atatgcaaag cagaaaccct cgtgcaggcc cgaaagagaa agcgaaccag
     4321 tatcgagaac cgagtgagag gcaacctgga gaatttgttc ctgcagtgcc cgaaacccac
     4381 actgcagcag atcagccaca tcgcccagca gcttgggctc gagaaggatg tggtccgagt
     4441 gtggttctgt aaccggcgcc agaagggcaa gcgatcaagc agcgactatg cacaacgaga
     4501 ggattttgag gctgctgggt ctcctttctc agggggacca gtgtcctttc ctctggcccc
     4561 agggccccat tttggtaccc caggctatgg gagccctcac ttcactgcac tgtactcctc
     4621 ggtccctttc cctgaggggg aagcctttcc ccctgtctcc gtcaccactc tgggctctcc
     4681 catgcattca aactgaggtg cctgccctct agaatcgaat tcgaagcttg agctcgagat
     4741 ctgagtccgg tagcgctagt tcgaaggatc cgcatgcatc tagggcggcc aattccgccc
     4801 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     4861 gtttgtctat atgtgatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     4921 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     4981 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     5041 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     5101 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     5161 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     5221 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     5281 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     5341 cgtggttttc ctttgaaaaa cacgatgata agcttgccac aacccacaag gagacgacct
     5401 tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc
     5461 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg
     5521 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     5581 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     5641 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     5701 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     5761 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     5821 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     5881 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     5941 gccgacgtcg agtgcccgaa ggaccgcgcg acctggtgca tgacccgcaa gcccggtgcc
     6001 tgataatagg cggccgctcg agacctagaa aaacatggag caatcacaag tagcaataca
     6061 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     6121 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     6181 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     6241 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     6301 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     6361 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     6421 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     6481 gcaaagaaat gaatatcaga gagtgagagg ccttgacatt ataatagatt tagcaggaat
     6541 tgaactagga gtggagcaca caggcaaagt tctagagctc gctgatcagc ctcgactgtg
     6601 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     6661 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     6721 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     6781 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     6841 agctgggggc gcgcccctcg aggccgccat ggtcatagct gtttgacgtc aggtggcact
     6901 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6961 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     7021 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     7081 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     7141 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     7201 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     7261 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7321 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     7381 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     7441 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     7501 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7561 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7621 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7681 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7741 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7801 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7861 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7921 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7981 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     8041 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     8101 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     8161 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     8221 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     8281 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     8341 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     8401 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
     8461 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8521 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8581 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8641 aacgccagca acgcggcctt tttacggttc ctggcctttt gc
//
