LOCUS       V034432                 8998 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034432
VERSION     V034432
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             2302..3852
                     /label="ALDH2(NM_000690)"
                     /note="ALDH2(NM_000690)"
                     /gene="ALDH2"
     CDS             3859..3924
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3968..4179
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4192..4460
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4487..5086
                     /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    5093..5681
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5755..5988
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6060..6181
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6200..6335
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6373..6389)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6397..6413
                     /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(6421..6451)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6466..6487
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6775..7363)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7534..8394)
                     /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(8395..8499)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8973..8989
                     /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgttgcgc gctgccgccc gcttcgggcc ccgcctgggc
     2341 cgccgcctct tgtcagccgc cgccacccag gccgtgcctg cccccaacca gcagcccgag
     2401 gtcttctgca accagatttt cataaacaat gaatggcacg atgccgtcag caggaaaaca
     2461 ttccccaccg tcaatccgtc cactggagag gtcatctgtc aggtagctga aggggacaag
     2521 gaagatgtgg acaaggcagt gaaggccgcc cgggccgcct tccagctggg ctcaccttgg
     2581 cgccgcatgg acgcatcaca caggggccgg ctgctgaacc gcctggccga tctgatcgag
     2641 cgggaccgga cctacctggc ggccttggag accctggaca atggcaagcc ctatgtcatc
     2701 tcctacctgg tggatttgga catggtcctc aaatgtctcc ggtattatgc cggctgggct
     2761 gataagtacc acgggaaaac catccccatt gacggagact tcttcagcta cacacgccat
     2821 gaacctgtgg gggtgtgcgg gcagatcatt ccgtggaatt tcccgctcct gatgcaagca
     2881 tggaagctgg gcccagcctt ggcaactgga aacgtggttg tgatgaaggt agctgagcag
     2941 acacccctca ccgccctcta tgtggccaac ctgatcaagg aggctggctt tccccctggt
     3001 gtggtcaaca ttgtgcctgg atttggcccc acggctgggg ccgccattgc ctcccatgag
     3061 gatgtggaca aagtggcatt cacaggctcc actgagattg gccgcgtaat ccaggttgct
     3121 gctgggagca gcaacctcaa gagagtgacc ttggagctgg gggggaagag ccccaacatc
     3181 atcatgtcag atgccgatat ggattgggcc gtggaacagg cccacttcgc cctgttcttc
     3241 aaccagggcc agtgctgctg tgccggctcc cggaccttcg tgcaggagga catctatgat
     3301 gagtttgtgg agcggagcgt tgcccgggcc aagtctcggg tggtcgggaa cccctttgat
     3361 agcaagaccg agcaggggcc gcaggtggat gaaactcagt ttaagaagat cctcggctac
     3421 atcaacacgg ggaagcaaga gggggcgaag ctgctgtgtg gtgggggcat tgctgctgac
     3481 cgtggttact tcatccagcc cactgtgttt ggagatgtgc aggatggcat gaccatcgcc
     3541 aaggaggaga tcttcgggcc agtgatgcag atcctgaagt tcaagaccat agaggaggtt
     3601 gttgggagag ccaacaattc cacgtacggg ctggccgcag ctgtcttcac aaaggatttg
     3661 gacaaggcca attacctgtc ccaggccctc caggcgggca ctgtgtgggt caactgctat
     3721 gatgtgtttg gagcccagtc accctttggt ggctacaaga tgtcggggag tggccgggag
     3781 ttgggcgagt acgggctgca ggcatacact gaagtgaaaa ctgtcacagt caaagtgcct
     3841 cagaagaact caggatccga ctacaaagac catgacggtg attataaaga tcatgacatc
     3901 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc
     3961 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     4021 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     4081 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     4141 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
     4201 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
     4261 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
     4321 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
     4381 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
     4441 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga ccgagtacaa
     4501 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     4561 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg
     4621 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     4681 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     4741 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     4801 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     4861 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     4921 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     4981 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5041 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg acaatcaacc
     5101 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5161 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5221 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5281 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5341 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5401 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5461 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     5521 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5581 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     5641 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa
     5701 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     5761 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt
     5821 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     5881 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     5941 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     6001 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     6061 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6121 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6181 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     6241 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6301 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa
     6361 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6421 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6481 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6541 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6601 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6661 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6721 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6781 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6841 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6901 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6961 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7021 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7081 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7141 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7201 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7261 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7321 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7381 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7441 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7501 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7561 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7621 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7681 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7741 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7801 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7861 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7921 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7981 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8041 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8101 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8161 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8221 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8281 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8341 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8401 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8461 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8521 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     8581 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     8641 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     8701 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     8761 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     8821 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     8881 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     8941 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
