LOCUS       V047567                 9340 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047567
VERSION     V047567
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..4194
                     /label="Gbp4(NM_001256005)"
                     /note="Gbp4(NM_001256005)"
                     /gene="Gbp4"
     CDS             4201..4266
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4310..4521
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4534..4802
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4829..5428
                     /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    5435..6023
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6097..6330
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6402..6523
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6542..6677
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6715..6731)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6739..6755
                     /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(6763..6793)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6808..6829
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7117..7705)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7876..8736)
                     /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(8737..8841)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9315..9331
                     /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 catgacccaa ccacaaatgg ctcccatttg tcttgtggaa
     2341 aaccacaatg aacagctgtc agtgaaccag gaagccatag agattctgga caagatttct
     2401 cagccagtgg tagtagtggc tattgttgga tggtcccata cagggaagtc ctatttgatg
     2461 aactgtttgg cgggacagaa tcacgtgagt ggcacgctgc ctaccagcca aaggtttccc
     2521 tctgggctcc accgtgcagt ctcagaccaa gggcatctgg atgtggtgca tgccccaccc
     2581 caccaagcca gagcactggt cctcctggac accgagggcc tgggcgatgt ggaaaagggt
     2641 gatcctaaga acgacttgtg gatctttgcc ctcagtgtgc ttctgagcag caccttcgtc
     2701 tacaacagca tgaacaccat caaccaccaa gccctggagc agctgcatta tgtcacagaa
     2761 ctcactgagc tgatcagagc aaaatcttcc ccaaatcccc atggaataaa gaattccaca
     2821 gagtttgtga gtttctttcc agactttgtc tggactgttc gggatttcat gctggagctg
     2881 aagttaaatg gggaagacat cacaagtgat gagtacctgg agaatgccct gaagctgatc
     2941 ccaggtaaca atcccagaat ccaagcatcc aattcggcca gggaatgcat cagacgtttc
     3001 tttcctaatc ggaagtgttt tgtctttgaa tggccaactc atgacataga actcataaaa
     3061 caacttgaga ctatttcaga agaccaattg gatcctacgt ttaaggaaag tgcaatggct
     3121 tttgcttctt atatcttcac ttatgccaag atcaagaccc tcagagaggg aattaaggtc
     3181 actgggaatg gactgggtac tctggtgaca acctacgtgg atgccatcaa cagtggagca
     3241 gtgccttgtc tggatgatgc tgtgacaact ctggcccagc gtgagaactc agtagctgtg
     3301 cagaaggcag ccagccacta cagtgagcag atggcccagc gactgagcct tcccacagac
     3361 acgatccagg agctgctgga tgtacatgca gcctgcgaga aggaagccat ggctgtcttt
     3421 atggagcatt ccttcaaaga cgaaaatcag caattcctga agaagctggt ggaattacta
     3481 cgtgagaaga atgggctttt cctgttgaag aatgaagagg catctgataa atactgccag
     3541 gaagaactgg atcgactttc aaaggatttg atggacaata tctcaacatt ttctgttcct
     3601 gggggacaca ggctctacat ggacatgagg gaaaagattg aacatgacta ctggcaggtt
     3661 cccaggaaag gggtgaaggc aagtgaagtc ttccagaact ttctgcagtc acaggccatc
     3721 atcgagagtt ccatcttgca ggcagataca gccctcactg ctgggcagaa ggccattgca
     3781 gagaagcaca ccaagaagga ggcagctgag aaggagcagg atctgctaag acagaagcag
     3841 aaggagcatc aggagtatat ggaggctcaa gagaaaagga acaaggaaaa cctagagcaa
     3901 ctgagaagga agctggagca ggagagagag cagctcatca aagaccataa catgatgctg
     3961 gagaagctaa cgaaggaaca aaagactttc cgtgaggaag gatataagac acaagctgag
     4021 gaattgcgtc gagagatcca tcaactggga cataacatca aggaaatgaa acaaaatggt
     4081 gattcccttg tggaaagtat tttaagaagt tggttttcat ttatttcacc cccttcagag
     4141 tcagaaaagg ctataagcag cgttttatca ctccttagga aaaaggatag actgggatcc
     4201 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     4261 gacaagtagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     4321 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     4381 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4441 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4501 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     4561 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     4621 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     4681 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4741 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4801 tccaagctgt gaccggcgcc tacgctagat gaccgagtac aagcccacgg tgcgcctcgc
     4861 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4921 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4981 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5041 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5101 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5161 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5221 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5281 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5341 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5401 gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg
     5461 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5521 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5581 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5641 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5701 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5761 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5821 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     5881 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5941 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6001 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     6061 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6121 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6181 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6241 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6301 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6361 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6421 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6481 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     6541 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6601 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6661 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     6721 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6781 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6841 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6901 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6961 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7021 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7081 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7141 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7201 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7261 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7321 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7381 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7441 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7501 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7561 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7621 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7681 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7741 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7801 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7861 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7921 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7981 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8041 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8101 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8161 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8221 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8281 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8341 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8401 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8461 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8521 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8581 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8641 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8701 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8761 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8821 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     8881 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     8941 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     9001 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     9061 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     9121 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     9181 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9241 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9301 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
