LOCUS       V026821                 9798 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026821
VERSION     V026821
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2301..4652
                     /label="Usp1(NM_146144)"
                     /note="Usp1(NM_146144)"
                     /gene="Usp1"
     CDS             4659..4724
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4768..4979
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4992..5260
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5287..5886
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5893..6481
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6555..6788
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6860..6981
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7000..7135
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7173..7189)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7197..7213
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7221..7251)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7266..7287
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7575..8163)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8334..9194)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(9195..9299)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9773..9789
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgcctggcg tcatacctag tgaaagtaat gggctttcaa
     2341 gaggcagtcc atccaagaaa aacagactgt ctctgaagtt ttttcagaaa aaggaaacta
     2401 aaagagcctt agactttact gattctcaag aaaatgaaga aaaaacttct gaatatagag
     2461 gatctgagat tgaccaagtt gttcctgctg cacagtcctc cccagtaagc tgcgagaaga
     2521 gagagaactt gttaccattt gtgggattga ataaccttgg caacacttgt tatctgaata
     2581 gtattcttca ggtgttatac ttttgtcctg gctttaagac tggagtgaag cacttattta
     2641 atattatttc aaggaagaaa gaagctctga aggatgactc taatcagaag gataagggaa
     2701 gctgcaaaga agagtctttg gcaagttatg agcttatatg tagtttacag tccttaataa
     2761 tttcagttga acagcttcag gctagttttc tcttaaatcc agagaaatat acagatgaac
     2821 ttgctacaca gccaaggcga ctgctgaaca cactcaggga actcaaccct atgtatgaag
     2881 gatttctaca gcatgatgca caggaagtgc tacagtgtat tctgggaaac attcaagaaa
     2941 catgccaact cctaaaaaaa gaagaaataa aaaacctggc tgaattatcc ggcaaggttg
     3001 aagaacaatc tcttcagaaa gaggaaacag gtggaattac cagcacagag attgacagta
     3061 tgaggaatac ggaggacgtc aaagagcagc tccccaaagg gaactggaaa agaaaaagtg
     3121 acagtgagtc gagtaacgtg aagaaaaagg ttaaactgtc cagggaatcc cagccattgg
     3181 aagaaaacca gagacaaacc agatccaaga gaaaagctat cggagacaca ctagaggctg
     3241 ctcctaagat catccccaag tgtgtttctg aaagtgagag tgcaaagccc tcacagaaga
     3301 aatcaaaagt taaaataaat tggttaaagc catccactaa gcaacccagc attctttcca
     3361 agttctgtag tcttggaaaa ataacaacta accagcgctc caaaggacaa cctaaggaaa
     3421 aggagggtga tgttgaagag gacttggaga agtacggaag tgaccacaca gctaatggtg
     3481 gacccgaatc tccaggaagt agcgtcacac ctgtggacag cagtgaagcc aagtccggaa
     3541 acaaaggtgc tgaacagatt ggctttgagc tagtagagaa actgtttcaa ggtcagctcg
     3601 tgttaaggac tcggtgtttg gaatgtgaaa gcttaacaga aagaagagaa gatttccagg
     3661 atattagtgt tccagtgcaa gaagatgagc tttccaaggt cgaggagagc tctgaaattt
     3721 ctccagagcc aaaaacagaa atgaagacct tgagatgggc aatttcacag tttgcttcag
     3781 tggagagaat tgtaggagag gataaatact tctgtgaaaa ttgccatcat tatacggaag
     3841 cagagcgaag tctcctgttt gataaaatgc ctgaagttat cactattcat ctgaagtgct
     3901 ttgctgctag tggcttggag tttgactgtt acggtggtgg actttccaag atcaacactc
     3961 ctttattgac acctcttaaa ctgtcactag aagaatggag cacaaagccg acgaatgaca
     4021 gctatggatt atttgccgtt gtgatgcaca gtggcattac tattagtagt gggcactata
     4081 ccgcatctgt taaagtcact gaccttaaca gtctagagct cgatgagggg aattttgtgg
     4141 ttgaccagat gtgtgaacta ggtaagccag agccactgac cgaggagcaa gcaaggggga
     4201 cggctgggaa ctacgacgat gaagtatcaa ttagagttgg tggaaatgcc cagccaagta
     4261 aagttttgaa caaaaaaaat gtagaaggta ttggacttct cggtggacaa aagagcaaag
     4321 cagattatga gctgtacaac aaagcatcta atcctgataa ggtggttggg acaccattca
     4381 ccgacaatag aaattctgaa actaatgata ctactaatgg gacccacgaa tctgacagga
     4441 acaaggaatc cagtgaccaa acaggcgtta acatgaacgg acttgagaac aaaatttcct
     4501 atgtagtgca aagcttaaag gagtacgagg ggaagtggtt gctctttgac gattctgagg
     4561 taaaagtgac agaagagaaa gactttctga attctctctc cccttctaca tcccctacgt
     4621 ctactcccta cttgctattc tataaaaaat taggatccga ctacaaagac catgacggtg
     4681 attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg ccgcgaagga
     4741 tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     4801 agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact
     4861 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     4921 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc
     4981 tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc
     5041 atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt
     5101 ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt
     5161 ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct
     5221 acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta
     5281 cgctagatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg
     5341 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac
     5401 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     5461 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     5521 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     5581 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     5641 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     5701 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     5761 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     5821 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     5881 gcctgagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5941 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6001 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6061 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6121 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6181 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6241 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     6301 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6361 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6421 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6481 cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6541 agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc
     6601 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6661 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6721 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6781 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     6841 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     6901 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6961 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     7021 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     7081 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac
     7141 ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     7201 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     7261 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7321 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7381 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7441 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7501 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7561 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7621 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7681 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7741 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7801 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7861 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7921 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7981 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     8041 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     8101 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     8161 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     8221 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     8281 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     8341 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8401 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8461 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8521 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8581 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8641 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8701 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8761 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8821 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8881 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8941 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     9001 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     9061 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     9121 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     9181 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     9241 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     9301 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     9361 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     9421 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     9481 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     9541 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     9601 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     9661 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     9721 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     9781 acggccagtg ccaagctg
//
