LOCUS       V034403                 9373 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034403
VERSION     V034403
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..4227
                     /label="Dlat(NM_145614)"
                     /note="Dlat(NM_145614)"
                     /gene="Dlat"
     CDS             4234..4299
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4343..4554
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4567..4835
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4862..5461
                     /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    5468..6056
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6130..6363
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6435..6556
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6575..6710
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6748..6764)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6772..6788
                     /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(6796..6826)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6841..6862
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7150..7738)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7909..8769)
                     /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(8770..8874)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9348..9364
                     /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 catgtggcgc gtctgtgcgc ggagggcgcg gagtgctgtg
     2341 cccagggatg ggttcagggc tcggtgggcg gccttgaagg aaggccccgg ggccccatgc
     2401 gggagcccgc ggattggccc cgctgcggtc cgttgcggta gtgggatccc ccgctatggg
     2461 gtccgctcct tgtgcggctg gagctccggc tccggcactg tcccgcgtaa ccgcttgctg
     2521 cgacaactcc tggggtcgcc cagccgccgc tcttacagcc tccctccgca tcagaaggtt
     2581 ccattacctt ccctttcccc tacaatgcag gcaggcacca tagcccgctg ggaaaaaaaa
     2641 gaaggggaaa aaatcagtga gggtgatcta attgcagagg ttgaaactga taaagccact
     2701 gttgggtttg agagcctgga ggagtgctat atggccaaga tccttgttcc tgaagggacc
     2761 agagatgttc cagttgggtc catcatctgt atcacagttg aaaagcctca ggatattgag
     2821 gcctttaaaa attatacatt ggatttggcg gcggcagctg ccccacaggc agccccagct
     2881 gcagccccag ctccagctgc tgccccagcg gcaccttctg caagcgctcc cggtagctcc
     2941 tatcctactc atatgcagat tgttcttcct gccctctccc caaccatgac catgggcacg
     3001 gtccagaggt gggaaaagaa agtgggcgag aagctgagtg aaggagactt gctggcagag
     3061 atagagaccg acaaggccac cataggcttt gaagtacaag aagaaggtta tctggcaaaa
     3121 atcctggtcc ctgaaggcac aagggatgtt cctctgggag ctccactatg tatcatagta
     3181 gaaaaacagg aagatatagc agcatttgca gactacaggc caacagaagt gaccagctta
     3241 aagccacagg cagcaccacc tgccccaccc ccggtggcag ctgttcctcc cactccccag
     3301 cctgtagcac ctaccccctc agctgctcct gctggaccaa agggaagggt gttcgttagt
     3361 cctcttgcga agaagttggc agcagagaaa gggattgacc tcacccaagt taaagggacg
     3421 ggaccagaag gcagaatcat caagaaggac attgactctt ttgtgccttc taaggctgct
     3481 cctgctgcag cagctgccat ggctcccccg ggtccaagag tggcaccagc tcctgcaggt
     3541 gtcttcacag acatccccat cagcaacatt cgtcgagtga ttgcgcagag gctcatgcag
     3601 tcgaagcaga ctatacctca ttattacctt tctgtcgatg taaatatggg agaggtgctg
     3661 ttggtacgga aggaacttaa taagatgctt gaagggaaag ggaaaatctc cgtcaacgac
     3721 ttcattataa aggcttcagc tttggcctgt ctgaaagttc ccgaagcaaa ctcgtcttgg
     3781 atggacacag ttataaggca aaatcatgtg gttgacgtca gtgttgctgt cagtacccct
     3841 gcaggactta tcacccctat tgtgtttaat gcacacataa aaggactgga aaccattgct
     3901 agtgatgttg tttctttagc ctccaaagcg agagagggta aacttcagcc tcatgagttc
     3961 cagggtggaa catttacaat ctccaactta gggatgtttg gaattaagaa tttctctgca
     4021 attattaatc cacctcaggc atgtattctg gcaatcggtg cttcagagga taaactgatc
     4081 ccagcagata atgagaaagg ctttgatgtg gctagtgtga tgtccgttac actcagctgt
     4141 gatcatcgag ttgtggatgg agcagttgga gcccagtggc ttgctgaatt caaaaagtac
     4201 cttgaaaagc ctataactat gttgttagga tccgactaca aagaccatga cggtgattat
     4261 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc
     4321 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     4381 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa
     4441 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     4501 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag
     4561 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca
     4621 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc
     4681 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc
     4741 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc
     4801 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta
     4861 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
     4921 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
     4981 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     5041 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     5101 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     5161 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     5221 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     5281 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     5341 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     5401 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     5461 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5521 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5581 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5641 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5701 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5761 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5821 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     5881 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5941 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     6001 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     6061 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6121 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta
     6181 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6241 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6301 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6361 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6421 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6481 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6541 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc
     6601 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6661 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg
     6721 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6781 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6841 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6901 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6961 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7021 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7081 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7501 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7561 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7681 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7741 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7801 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7861 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7921 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7981 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8041 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8101 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8161 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8221 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8281 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8341 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8401 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8461 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8521 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8581 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8641 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8701 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8761 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8821 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8881 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     8941 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     9001 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     9061 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     9121 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     9181 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     9241 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     9301 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     9361 cagtgccaag ctg
//
