LOCUS       V026883                10280 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026883
VERSION     V026883
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     promoter        2337..3519
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             3548..4927
                     /label="Gipr(NM_001080815)"
                     /note="Gipr(NM_001080815)"
                     /gene="Gipr"
     CDS             4937..5002
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5030..5616
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5636..6235
                     /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    6249..6837
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7044..7677
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7805..7821)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7829..7845
                     /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(7853..7883)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7898..7919
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8207..8795)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8966..9826)
                     /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(9827..9931)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9979..10113
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
     2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
     2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
     2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtacgaatt
     3541 cgccaccatg cccctgcggt tgctgcttct gctgctgtgg ttgtggggac tccagtgggc
     3601 ggagacagac tctgaggggc agaccaccac gggggagctg taccagcgct gggagcacta
     3661 cggccaggag tgccaaaaga tgttggagac cacagaacct ccctcaggcc tggcctgtaa
     3721 cggttccttc gatatgtatg cctgctggaa ctacacggcc gccaacacca ctgctcgggt
     3781 gtcttgcccc tggtatctgc cctggttccg tcaggtgtct gcaggctttg tcttccgcca
     3841 gtgtggcagt gatggccagt ggggatcttg gagagaccac actcagtgtg agaatccaga
     3901 gaagaatggg gcttttcagg accagacgct gatcctggag cgcctgcaga tcatgtatac
     3961 cgtgggctac tccctgtccc tgacgactct gctgctagcc ctactcatct taagtttgtt
     4021 caggcggctg cactgcactc gtaattacat tcacatgaac ctgttcacgt ctttcatgct
     4081 gcgggcagca gccatcctca cccgagatca gctgctgcct ccactgggtc cctacactgg
     4141 agaccaggcc cctaccccgt ggaaccaggc cctagctgcc tgccgcacgg cccagatcat
     4201 gacccaatat tgtgtgggag ccaattacac ctggctactg gtggagggtg tttatctgca
     4261 ccatctgctg gtgatcgtgg gacgctcaga aaagggccac ttccgctgct acctgcttct
     4321 tggctggggg gcccccgcgc ttttcgtcat tccctgggtg atcgtcaggt acctgcgcga
     4381 gaacacacag tgctgggagc gcaacgaagt caaagccatt tggtggatca ttcgcactcc
     4441 catcctaata accatcttga tcaatttcct catcttcatc cgcatccttg gcatccttgt
     4501 ttcaaagctg aggacacggc agatgcgctg ccccgactac cgactaaggc tggctcgctc
     4561 cacgctgaca ctggtgcccc tgctgggtgt ccacgaggtg gtgtttgcgc ctgtgacgga
     4621 ggaacaggtt gaaggctccc tgcgcttcgc caaactggcc tttgaaatct tcctaagttc
     4681 cttccagggt ttcctggtga gcgtgctcta ctgcttcatc aacaaagagg tgcagtcgga
     4741 gatccgccag ggttggcgcc accgccgcct gcgtctcagt cttcaagagc agcgtccacg
     4801 tccgcaccag gaactcgccc cccgggctgt gcctttgagc tctgcgtgcc gagaagctgc
     4861 cgttggcaac gccttgccct ctgggatgct gcatgtgcct ggggatgagg tcttggaaag
     4921 ttactgcagc ggccgcgact acaaagacca tgacggtgat tataaagatc atgacatcga
     4981 ctacaaggat gacgatgaca agtaggcctg cgatatccag cggatcccgc ccctctccct
     5041 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     5101 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     5161 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     5221 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     5281 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     5341 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     5401 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     5461 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     5521 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     5581 tcctttgaaa aacacgatga taagcttgcc acaacccaca aggagacgac cttccatgac
     5641 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac
     5701 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca
     5761 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     5821 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     5881 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     5941 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     6001 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     6061 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     6121 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     6181 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctagacgcg
     6241 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     6301 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     6361 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     6421 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     6481 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     6541 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6601 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     6661 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     6721 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     6781 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     6841 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     6901 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     6961 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     7021 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat
     7081 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc
     7141 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     7201 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
     7261 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct
     7321 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg
     7381 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
     7441 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt
     7501 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     7561 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     7621 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     7681 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     7741 gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc ttggcgtaat
     7801 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7861 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7921 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7981 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     8041 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     8101 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     8161 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     8221 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     8281 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     8341 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     8401 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     8461 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     8521 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     8581 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8641 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8701 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     8761 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8821 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8881 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8941 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     9001 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     9061 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     9121 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     9181 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     9241 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     9301 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     9361 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     9421 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     9481 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     9541 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9601 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9661 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9721 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9781 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9841 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9901 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     9961 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    10021 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    10081 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
    10141 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
    10201 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
    10261 tgtactacaa acttagtagt
//
