LOCUS       Exported                9378 bp ds-DNA     circular SYN 01-AUG-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSin-EF2-Nanog-Pur
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9378)
  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 Nov 20. ():.
  PUBMED    18029452
REFERENCE   2  (bases 1 to 9378)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 1, 2019 from SnapGene 4.1.8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9378
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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."
     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"
     regulatory      3610..3619
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    3616..4299
                     /label=ECFP
     CDS             4699..5409
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     LCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLEF
                     LTAAGITEGMNELYK"
     misc_feature    5494..6067
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5661..5678)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5888..5907
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6100..6699
                     /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(6100..6119)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6596..6616
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6902..7135
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7286..7303)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7292..7516
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      7559..7568
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        7612..7716
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7717..8577
                     /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(7935..7954)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8748..9336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9237..9256
                     /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 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3661 ctggtcgagc tggacggcga cgtaaacggc cacaggttca gcgtgtccgg cgagggcgag
     3721 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3781 gtgccctggc ccaccctcgt gaccaccctg acctggggcg tgcagtgctt cagccgctac
     3841 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3901 gagcgtacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3961 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4021 aacatcctgg ggcacaagct ggagtacaac tacatcagcc acaacgtcta tatcaccgcc
     4081 gacaagcaga agaacggcat caaggcccac ttcaagatcc gccacaacat cgaggacggc
     4141 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4201 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     4261 cgcgatcaca tggtcctgct ggagttcgtg accgccgccc gcatgcattg gtattttggg
     4321 aagatcactc gtcgggagtc cgagcggctg ctgctcaacc ccgaaaaccc ccggggaacc
     4381 ttcttggtcc gggagagcga gacgacaaaa ggtgcctatg ccctctccgt ttctgacttt
     4441 gacaacgcca aggggctcaa tgtgaagcac tacaagatcc gcaagctgga cagcggcggc
     4501 ttctacatca cctcacgcac acagttcagc agcctgcagc agctggtggc ctactactcc
     4561 aaacatgctg atggcttgtg ccaccgcctg actaacgtct gtgggtctac atctggatct
     4621 gggaagccgg gttctggtga gggttctatg gacacgagcg tgttcgagag cccctacagc
     4681 gacccagagg aggagctctc taaaggtgaa gaattattca ctggtgttgt cccaattttg
     4741 gttgaattag atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt
     4801 gatgctacgt acggtaaatt gaccttaaaa ttactctgta ctactggtaa attgccagtt
     4861 ccatggccaa ccttagtcac tactttaggt tatggtgttc aatgttttgc tagataccca
     4921 gatcatatga aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa
     4981 agaactattt ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa
     5041 ggtgatacct tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac
     5101 attttaggtc acaaattgga atacaactat aactctcaca atgtttacat cactgctgac
     5161 aaacaaaaga atggtatcaa agctaacttc aaaattagac acaacattga agatggtggt
     5221 gttcaattag ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta
     5281 ccagacaacc attacttatc ctatcaatct gccttattca aagatccaaa cgaaaagaga
     5341 gaccacatgg tcttgttaga atttttgact gctgctggta ttaccgaggg tatgaatgaa
     5401 ttgtacaaat aagaattcga agcttgagct cgagatctga gtccggtagc gctagttcga
     5461 aggatccgca tgcatctagg gcggccaatt ccgcccctct ccctcccccc cccctaacgt
     5521 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt gattttccac
     5581 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     5641 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     5701 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     5761 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     5821 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     5881 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     5941 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     6001 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     6061 atgataagct tgccacaacc cacaaggaga cgaccttcca tgaccgagta caagcccacg
     6121 gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc
     6181 gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc
     6241 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     6301 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     6361 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     6421 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     6481 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     6541 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     6601 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgagtg cccgaaggac
     6661 cgcgcgacct ggtgcatgac ccgcaagccc ggtgcctgat aataggcggc cgctcgagac
     6721 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
     6781 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
     6841 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     6901 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     6961 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7021 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7081 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     7141 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     7201 gagaggcctt gacattataa tagatttagc aggaattgaa ctaggagtgg agcacacagg
     7261 caaagttcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     7321 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     7381 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     7441 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     7501 gcggtgggct ctatggcttc tgaggcggaa agaaccagct gggggcgcgc ccctcgaggc
     7561 cgccatggtc atagctgttt gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     7621 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7681 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7741 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7801 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7861 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7921 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7981 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     8041 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     8101 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     8161 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     8221 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     8281 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     8341 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     8401 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     8461 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     8521 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8581 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8641 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8701 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8761 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8821 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8881 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8941 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     9001 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     9061 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     9121 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     9181 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     9241 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     9301 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     9361 cggttcctgg ccttttgc
//
