LOCUS       sequence-9806-ex        9030 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 9030)
  AUTHORS   Yu J, Hu K, Smuga-Otto K, Tian S, Stewart R, Slukvin II, Thomson JA
  TITLE     Human Induced Pluripotent Stem Cells Free of Vector and Transgene 
            Sequences
  JOURNAL   Science. 2009 Mar 26.
  PUBMED    19325077
REFERENCE   2  (bases 1 to 9030)
  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. 
            2009 Mar 26."
COMMENT     Sequence Label: pSIN4-EF2-O2S
FEATURES             Location/Qualifiers
     source          1..9030
                     /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    4766..5339
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4789..5341
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4933..4950)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5160..5179
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             6550..6783
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6938..6955)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6944..7168
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      7211..7220
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        7264..7368
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7369..7437,7438..8229)
                     /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(7587..7606)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8400..8988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8889..8908
                     /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 ngagtcgggg 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 agtttacgcg tgggggcggc cgctttctcg
     4741 aggccatcga tgtttaaacg atccgcccct ctccctcccc cccccctaac gttactggcc
     4801 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     4861 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     4921 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     4981 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     5041 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     5101 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     5161 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     5221 tgggatctga tctggggcct cggtacacat gctttacatg tgtttagtcg aggttaaaaa
     5281 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     5341 atggccacac cgcgatcgcg aattcgattc cgcatgtaca acatgatgga gacggagctg
     5401 aagccgccgg gcccgcagca aacttcgggg ggcggcggcg gcaactccac cgcggcggcg
     5461 gccggcggca accagaaaaa cagcccggac cgcgtcaagc ggcccatgaa tgccttcatg
     5521 gtgtggtccc gcgggcagcg gcgcaagatg gcccaggaga accccaagat gcacaactcg
     5581 gagatcagca agcgcctggg cgccgagtgg aaacttttgt cggagacgga gaagcggccg
     5641 ttcatcgacg aggctaagcg gctgcgagcg ctgcacatga aggagcaccc ggattataaa
     5701 taccggcccc ggcggaaaac caagacgctc atgaagaagg ataagtacac gctgcccggc
     5761 gggctgctgg cccccggcgg caatagcatg gcgagcgggg tcggggtggg cgccggcctg
     5821 ggcgcgggcg tgaaccagcg catggacagt tacgcgcaca tgaacggctg gagcaacggc
     5881 agctacagca tgatgcagga ccagctgggc tacccgcagc acccgggcct caatgcgcac
     5941 ggcgcagcgc agatgcagcc catgcaccgc tacgacgtga gcgccctgca gtacaactcc
     6001 atgaccagct cgcagaccta catgaacggc tcgcccacct acagcatgtc ctactcgcag
     6061 cagggcaccc ctggcatggc tcttggctcc atgggttcgg tggtcaagtc cgaggccagc
     6121 tccagccccc ctgtggttac ctcttcctcc cactccaggg cgccctgcca ggccggggac
     6181 ctccgggaca tgatcagcat gtatctcccc ggcgccgagg tgccggaacc cgccgccccc
     6241 agcagacttc acatgtccca gcactaccag agcggcccgg tgcccggcac ggccattaac
     6301 ggcacactgc ccctctcaca catgtgaggg ccggaaatca ctagctagtt cgaaggatcc
     6361 ctcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     6421 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
     6481 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6541 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     6601 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6661 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6721 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6781 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6841 cagagagtga gaggccttga cattataata gatttagcag gaattgaact aggagtggag
     6901 cacacaggca aagttctagc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     6961 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     7021 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     7081 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     7141 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctgggggcgc
     7201 gcccctcgag gccgccatgg tcatagctgt ttgacgtcag gtggcacttt tcggggaaat
     7261 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7321 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     7381 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7441 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7501 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7561 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     7621 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7681 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7741 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7801 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7861 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     7921 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7981 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     8041 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     8101 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8161 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8221 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8281 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8341 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     8521 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     8581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8761 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     8821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     9001 gcggcctttt tacggttcct ggccttttgc
//
