LOCUS       V043389                 9214 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043389
VERSION     V043389
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..4068
                     /label="Gbp2(NM_010260)"
                     /note="Gbp2(NM_010260)"
                     /gene="Gbp2"
     CDS             4075..4140
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4184..4395
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4408..4676
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4703..5302
                     /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    5309..5897
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5971..6204
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6276..6397
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6416..6551
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6589..6605)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6613..6629
                     /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(6637..6667)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6682..6703
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6991..7579)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7750..8610)
                     /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(8611..8715)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9189..9205
                     /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 catggcctca gagatccaca tgtcggaacc catgtgcctc
     2341 attgagaaca ctgaggctca actagtgatc aaccaggagg ctctgaggat cctgtctgcc
     2401 attacacagc ctgtggtggt ggtggcaatc gtgggcctct accgcacagg caaatcctac
     2461 ctgatgaaca agctagctgg gaagaggaca ggcttttccc tgggctccac tgtgcagtct
     2521 cacaccaagg gcatctggat gtggtgtgtg cctcacccca agaaggcagg gcaaaccctg
     2581 gttctgcttg acactgaggg ccttgaagat gttgagaagg gtgacaacca gaatgactgc
     2641 tggatctttg ctttggcagt cctcctcagc agcaccttca tctacaacag cataggaacc
     2701 atcaaccagc aggccatgga ccagctgcac tatgtgacgg agctaactga tcttatcaag
     2761 tcaaagtcat cacctgacca gagtggggta gacgattccg ctaactttgt gggcttcttt
     2821 ccaacctttg tgtggactct gagagatttc tccctcgagc tggaggttaa cggaaaaccc
     2881 gtcacttctg atgagtacct ggaacattcc ctgaccctga aaaaaggagc tgataagaaa
     2941 actaaaagct ttaatgagcc gcgactgtgc atcaggaaat tctttccaaa gaggaagtgc
     3001 ttcatctttg acaggcctgc tcagaggaag caacttagca aactggagac tctgcgggag
     3061 gaggagctgt gtggtgaatt tgtagaacaa gttgcagaat tcacctcata catcttgagc
     3121 tactcttctg tcaagactct gtgtggtggc atcatagtca atgggccacg tctaaagagc
     3181 ctggtgcaga cctatgttgg tgccatcagc aatgggtctc tcccctgcat ggagagtgct
     3241 gtgctgactt tggcccagat agagaactca gcagcagtgc aaaaggccat cacccactat
     3301 gaagaacaga tgaatcagaa gattcagatg cccacagaaa ccctccagga gctcctggat
     3361 ctgcacaggc caattgagag tgaggccatt gaggtcttcc tgaagaattc tttcaaggat
     3421 gtagaccaaa agttccagac agaattaggg aacctgctgg tagccaagcg agatgccttt
     3481 atcaagaaga acatggatgt ctcatcagct cgttgctcag acttgctgga ggatattttt
     3541 ggacccctgg aagaagaagt aaaattaggg acattttcta aaccaggagg ttactacctc
     3601 ttccttcaaa tgagacaaga gctagagaaa aagtataacc aggctcctgg gaaggggctc
     3661 caggcagaag cgatgctgaa aaactacttt gattccaagg cagatgttgt tgaaacactt
     3721 ctacagacgg atcagtcact cacagaggca gcaaaggagg tagaagagga acgtacgaag
     3781 gctgaagctg ctgaagctgc aaacagagag ttagaaaaga agcagaagga gttcgagctg
     3841 atgatgcagc agaaggaaaa gagttaccag gagcatgtga agaagctgac tgagaagatg
     3901 aaagacgaac agaaacagtt attagcagaa caggaaaaca tcatagctgc taaacttcgg
     3961 gaacaggaaa aatttcttaa ggaaggattc gagaatgaga gcaaaaaact tattcgagag
     4021 attgataccc tgaagcaaaa caaatcgtct gggaagtgca ctatactcgg atccgactac
     4081 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     4141 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     4201 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     4261 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     4321 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     4381 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     4441 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     4501 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     4561 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     4621 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag
     4681 ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4741 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4801 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4861 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4921 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4981 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5041 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5101 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5161 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5221 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5281 gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     5341 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5401 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5461 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5521 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5581 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5641 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5701 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5761 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5821 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5881 ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5941 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     6001 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6061 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6121 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6181 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6241 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6301 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6361 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6421 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6481 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6541 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     6601 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6661 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6721 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6781 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6841 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6901 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6961 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7021 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7081 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7141 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7201 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7261 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7321 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7381 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7441 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7501 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7561 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7621 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7681 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7741 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7801 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7861 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7921 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7981 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8041 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8101 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8161 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8221 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8281 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8341 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8401 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8461 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8521 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8581 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8641 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8701 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8761 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8821 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     8881 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     8941 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     9001 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     9061 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9121 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9181 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
