LOCUS       Exported                9669 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    FUW-SM
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9669)
  AUTHORS   Carey BW, Markoulaki S, Hanna J, Saha K, Gao Q, Mitalipova M, 
            Jaenisch R
  TITLE     Reprogramming of murine and human somatic cells using a single 
            polycistronic vector.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Jan 6. 106(1):157-62.
  PUBMED    19109433
REFERENCE   2  (bases 1 to 9669)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9669
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /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    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2622..2638
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2680..2713
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        2763..3974
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3811..3830
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4990..5055
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     protein_bind    6469..6502
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    6558..7146
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6611..6631)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7029..7040)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7149..7165)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7675..7855
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7917..8505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7997..8016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8676..9536)
                     /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     9299..9318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9537..9641)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccggg cccgctctag tcgaggtcga cggtatcgat
     2641 aagctcgctt cacgagattc cagcaggtcg agggacctaa taacttcgta tagcatacat
     2701 tatacgaagt tatattaagg gttccaagct taagcggcct aacccgtgtc ggctccagat
     2761 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc
     2821 tgccacgtca gacgaagggc gcagcgagcg tcctgatcct tccgcccgga cgctcaggac
     2881 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     2941 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     3001 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     3061 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     3121 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     3181 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     3241 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     3301 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     3361 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc
     3421 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tggggcacca tctggggacc
     3481 ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca
     3541 gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg
     3601 tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg
     3661 tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga
     3721 atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt
     3781 cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg
     3841 ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt
     3901 caatatgtaa ttttcagtgt tagactagta aattgtccgc taaattctgg ccgtttttgg
     3961 cttttttgtt agacgaagct tgggctgcag gtcgactcta gagaattcgc caccatgtat
     4021 aacatgatgg agacggagct gaagccgccg ggcccgcagc aagcttcggg gggcggcggc
     4081 ggaggaggca acgccacggc ggcggcgacc ggcggcaacc agaagaacag cccggaccgc
     4141 gtcaagaggc ccatgaacgc cttcatggta tggtcccggg ggcagcggcg taagatggcc
     4201 caggagaacc ccaagatgca caactcggag atcagcaagc gcctgggcgc ggagtggaaa
     4261 cttttgtccg agaccgagaa gcggccgttc atcgacgagg ccaagcggct gcgcgctctg
     4321 cacatgaagg agcacccgga ttataaatac cggccgcggc ggaaaaccaa gacgctcatg
     4381 aagaaggata agtacacgct tcccggaggc ttgctggccc ccggcgggaa cagcatggcg
     4441 agcggggttg gggtgggcgc cggcctgggt gcgggcgtga accagcgcat ggacagctac
     4501 gcgcacatga acggctggag caacggcagc tacagcatga tgcaggagca gctgggctac
     4561 ccgcagcacc cgggcctcaa cgctcacggc gcggcacaga tgcaaccgat gcaccgctac
     4621 gacgtcagcg ccctgcagta caactccatg accagctcgc agacctacat gaacggctcg
     4681 cccacctaca gcatgtccta ctcgcagcag ggcacccccg gtatggcgct gggctccatg
     4741 ggctctgtgg tcaagtccga ggccagctcc agcccccccg tggttacctc ttcctcccac
     4801 tccagggcgc cctgccaggc cggggacctc cgggacatga tcagcatgta cctccccggc
     4861 gccgaggtgc cggagcccgc tgcgcccagt agactgcaca tggcccagca ctaccagagc
     4921 ggcccggtgc ccggcacggc cattaacggc acactgcccc tgtcgcacat ggctagcggc
     4981 agcggcccgg tgaaacagac tttgaatttt gaccttctca agttggcggg agacgtcgag
     5041 tccaaccctg ggcccgcatg cctcgagggc gcgccactgg atttcctttg ggcgttggaa
     5101 accccgcaga cagccacgac gatgcccctc aacgtgaact tcaccaacag gaactatgac
     5161 ctcgactacg actccgtaca gccctatttc atctgcgacg aggaagagaa tttctatcac
     5221 cagcaacagc agagcgagct gcagccgccc gcgcccagtg aggatatctg gaagaaattc
     5281 gagctgcttc ccaccccgcc cctgtccccg agccgccgct ccgggctctg ctctccatcc
     5341 tatgttgcgg tcgctacgtc cttctcccca agggaagacg atgacggcgg cggtggcaac
     5401 ttctccaccg ccgatcagct ggagatgatg accgagttac ttggaggaga catggtgaac
     5461 cagagcttca tctgcgatcc tgacgacgag accttcatca agaacatcat catccaggac
     5521 tgtatgtgga gcggtttctc agccgctgcc aagctggtct cggagaagct ggcctcctac
     5581 caggctgcgc gcaaagacag caccagcctg agccccgccc gcgggcacag cgtctgctcc
     5641 acctccagcc tgtacctgca ggacctcacc gccgccgcgt ccgagtgcat tgacccctca
     5701 gtggtctttc cctacccgct caacgacagc agctcgccca aatcctgtac ctcgtccgat
     5761 tccacggcct tctctccttc ctcggactcg ctgctgtcct ccgagtcctc cccacgggcc
     5821 agccctgagc ccctagtgct gcatgaggag acaccgccca ccaccagcag cgactctgaa
     5881 gaagagcaag aagatgagga agaaattgat gtggtgtctg tggagaagag gcaaacccct
     5941 gccaagaggt cggagtcggg ctcatctcca tcccgaggcc acagcaaacc tccgcacagc
     6001 ccactggtcc tcaagaggtg ccacgtctcc actcaccagc acaactacgc cgcacccccc
     6061 tccacaagga aggactatcc agctgccaag agggccaagt tggacagtgg cagggtcctg
     6121 aagcagatca gcaacaaccg caagtgctcc agccccaggt cctcagacac ggaggaaaac
     6181 gacaagaggc ggacacacaa cgtcttggaa cgtcagagga ggaacgagct gaagcgcagc
     6241 ttttttgccc tgcgtgacca gatccctgaa ttggaaaaca acgaaaaggc ccccaaggta
     6301 gtgatcctca aaaaagccac cgcctacatc ctgtccattc aagcagacga gcacaagctc
     6361 acctctgaaa aggacttatt gaggaaacga cgagaacagt tgaaacacaa actcgaacag
     6421 cttcgaaact ctggtgcata aggcgcgccg aattcgtcga gggacctaat aacttcgtat
     6481 agcatacatt atacgaagtt atacatgttt aagggttccg gttccactag gtacaattcg
     6541 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     6601 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     6661 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     6721 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6781 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6841 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6901 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     6961 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     7021 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     7081 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     7141 ccccgcatcg ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt
     7201 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     7261 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     7321 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     7381 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag
     7441 aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta
     7501 gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta
     7561 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt
     7621 gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct
     7681 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7741 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7801 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg
     7861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     8341 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8401 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     8461 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8521 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8581 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8641 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8701 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8761 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8821 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8881 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8941 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     9001 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     9061 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     9121 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     9181 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     9241 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     9301 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     9361 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     9421 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     9481 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     9541 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     9601 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9661 ccacctgac
//
