LOCUS       sequence-8516-ex        8507 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8507)
  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 8507)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-Nanog-Pur
FEATURES             Location/Qualifiers
     source          1..8507
                     /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(4623..5197,5198..5200,5201..5209)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4790..4807)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5017..5036
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5229..5828
                     /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(5229..5248)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5725..5745
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6031..6264
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6415..6432)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6421..6645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      6688..6697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        6741..6845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6846..6914,6915..7706)
                     /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(7064..7083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7877..8465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8366..8385
                     /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 attcatgagt gtggatccag cttgtcccca aagcttgcct tgctttgaag
     3661 catccgactg taaagaatct tcacctatgc ctgtgatttg tgggcctgaa gaaaactatc
     3721 catccttgca aatgtcttct gctgagatgc ctcacacgga gactgtctct cctcttcctt
     3781 cctccatgga tctgcttatt caggacagcc ctgattcttc caccagtccc aaaggcaaac
     3841 aacccacttc tgcagagaag agtgtcgcaa aaaaggaaga caaggtcccg gtcaagaaac
     3901 agaagaccag aactgtgttc tcttccaccc agctgtgtgt actcaatgat agatttcaga
     3961 gacagaaata cctcagcctc cagcagatgc aagaactctc caacatcctg aacctcagct
     4021 acaaacaggt gaagacctgg ttccagaacc agagaatgaa atctaagagg tggcagaaaa
     4081 acaactggcc gaagaatagc aatggtgtga cgcagaaggc ctcagcacct acctacccca
     4141 gcctttactc ttcctaccac cagggatgcc tggtgaaccc gactgggaac cttccaatgt
     4201 ggagcaacca gacctggaac aattcaacct ggagcaacca gacccagaac atccagtcct
     4261 ggagcaacca ctcctggaac actcagacct ggtgcaccca atcctggaac aatcaggcct
     4321 ggaacagtcc cttctataac tgtggagagg aatctctgca gtcctgcatg cagttccagc
     4381 caaattctcc tgccagtgac ttggaggctg ccttggaagc tgctggggaa ggccttaatg
     4441 taatacagca gaccactagg tattttagta ctccacaaac catggattta ttcctaaact
     4501 actccatgaa catgcaacct gaagacgtgt gaagatgaat cgaattcgaa gcttgagctc
     4561 gagatctgag tccggtagcg ctagttcgaa ggatccgcat gcatctaggg cggccaattc
     4621 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     4681 tgtgcgtttg tctatatgtg attttccacc atattgccgt cttttggcaa tgtgagggcc
     4741 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     4801 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     4861 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     4921 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     4981 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     5041 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     5101 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     5161 ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc acaaggagac
     5221 gaccttccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc
     5281 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     5341 acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     5401 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     5461 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     5521 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     5581 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     5641 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     5701 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     5761 tcaccgccga cgtcgagtgc ccgaaggacc gcgcgacctg gtgcatgacc cgcaagcccg
     5821 gtgcctgata ataggcggcc gctcgagacc tagaaaaaca tggagcaatc acaagtagca
     5881 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     5941 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6001 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     6061 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6121 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6181 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6241 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6301 aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattataat agatttagca
     6361 ggaattgaac taggagtgga gcacacaggc aaagttctag agctcgctga tcagcctcga
     6421 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     6481 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     6541 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     6601 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     6661 gaaccagctg ggggcgcgcc cctcgaggcc gccatggtca tagctgtttg acgtcaggtg
     6721 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6781 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6841 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6901 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6961 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7021 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7081 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7141 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7201 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7261 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7321 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7381 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7441 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7501 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7561 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7621 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7681 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7741 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7801 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7861 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7921 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7981 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     8041 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8101 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8161 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8221 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
     8281 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8341 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8401 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8461 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgc
//
