LOCUS       Exported                8626 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSIN4-EF2-N2L
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8626)
  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 8626)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8626
                     /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."
     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    4683..5256
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4706..5258
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4850..4867)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5077..5096
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             6146..6379
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6534..6551)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6540..6764
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      6807..6816
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        6860..6964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6965..7825
                     /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(7183..7202)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7996..8584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8485..8504
                     /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 gcattcaaac tgaggtgcct
     4621 gccctctagt ttacgcgtgg gggcggccgc tttctcgagg ccatcgatgt ttaaacgatc
     4681 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     4741 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     4801 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     4861 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     4921 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     4981 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     5041 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     5101 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     5161 tacacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     5221 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaccgc gatcgcgaat
     5281 tcgattatcg ccatgggctc cgtgtccaac cagcagtttg caggtggctg cgccaaggcg
     5341 gcagaagagg cgcccgagga ggcgccggag gacgcggccc gggcggcgga cgagcctcag
     5401 ctgctgcacg gtgcgggcat ctgtaagtgg ttcaacgtgc gcatggggtt cggcttcctg
     5461 tccatgaccg cccgcgccgg ggtcgcgctc gaccccccag tggatgtctt tgtgcaccag
     5521 agtaagctgc acatggaagg gttccggagc ttgaaggagg gtgaggcagt ggagttcacc
     5581 tttaagaagt cagccaaggg tctggaatcc atccgtgtca ccggacctgg tggagtattc
     5641 tgtattggga gtgagaggcg gccaaaagga aagagcatgc agaagcgcag atcaaaagga
     5701 gacaggtgct acaactgtgg aggtctagat catcatgcca aggaatgcaa gctgccaccc
     5761 cagcccaaga agtgccactt ctgccagagc atcagccata tggtagcctc atgtccgctg
     5821 aaggcccagc agggccctag tgcacaggga aagccaacct actttcgaga ggaagaagaa
     5881 gaaatccaca gccctaccct gctcccggag gcacagaatt gagtttaaac naatcactag
     5941 ctagttcgaa ggatccctcg agacctagaa aaacatggag caatcacaag tagcaataca
     6001 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     6061 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     6121 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     6181 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     6241 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     6301 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     6361 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     6421 gcaaagaaat gaatatcaga gagtgagagg ccttgacatt ataatagatt tagcaggaat
     6481 tgaactagga gtggagcaca caggcaaagt tctagctaga gctcgctgat cagcctcgac
     6541 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     6601 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     6661 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     6721 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     6781 aaccagctgg gggcgcgccc ctcgaggccg ccatggtcat agctgtttga cgtcaggtgg
     6841 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6901 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6961 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7021 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7081 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7141 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7201 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7261 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7321 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7381 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7441 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7501 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7561 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7621 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7681 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7741 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7801 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7861 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7921 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7981 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     8041 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     8101 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     8161 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     8221 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     8281 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     8341 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc
     8401 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8461 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8521 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8581 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgc
//
