LOCUS       V022711                 9243 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022711
VERSION     V022711
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"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2815..3849
                     /label="GAS1(NM_002048)"
                     /note="GAS1(NM_002048)"
                     /gene="GAS1"
     CDS             3850..3915
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3946..4532
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             4552..5151
                     /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    5165..5753
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5960..6593
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6721..6737)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6745..6761
                     /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(6769..6799)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6814..6835
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7123..7711)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7882..8742)
                     /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(8743..8847)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8895..9029
                     /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 cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatggtg
     2821 gccgcgctgc tgggcggcgg cggcgaggcc cgcgggggga cagtgccggg cgcctggctg
     2881 tgcctgatgg cgctgctgca gctgctgggc tcggcgccgc ggggatcggg gctggcgcac
     2941 ggccgccgcc tcatctgctg gcaggcgctg ctgcagtgcc agggggagcc ggagtgcagc
     3001 tacgcctaca accagtacgc cgaggcgtgc gcgccggtgc tggcgcagca cggcgggggc
     3061 gacgcgcccg gggccgccgc cgccgctttc ccggcctcgg ccgcctcttt ctcgtcgcgc
     3121 tggcgctgcc cgagtcactg catctcggcc ctcattcagc tcaaccacac gcgccgcggg
     3181 cccgccctgg aggactgtga ctgcgcgcag gacgagaact gcaagtccac caagcgcgcc
     3241 attgagccgt gcctgccccg gacgagcggc ggcggcgcgg gcggccccgg cgcgggcggg
     3301 gtcatgggct gcaccgaggc ccggcggcgc tgcgaccgcg acagccgctg caacctggcg
     3361 ctgagccgct acctgaccta ctgcggcaaa gtcttcaacg ggctgcgctg cacggacgaa
     3421 tgccgcaccg tcattgagga catgctggct atgcccaagg cggcgctgct caacgactgc
     3481 gtgtgcgacg gcctcgagcg gcccatctgc gagtcggtca aggagaacat ggcccgcctg
     3541 tgcttcggcg ccgagctggg caacggcccc ggcagcagcg gctcggacgg gggcctggac
     3601 gactactacg atgaggacta cgatgacgag cagcgcaccg ggggcgcggg tggtgagcag
     3661 ccgctggacg acgacgacgg cgtcccgcac ccaccgcgcc cgggcagcgg cgctgctgca
     3721 tcgggcggcc gcggggacct gccctatggg cctgggcgca ggagcagcgg cggcggcggc
     3781 cgcttggcgc cccggggcgc ctggacccca ctcgcctcca tcttgctgct gctgcttggg
     3841 ccgctctttg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     3901 gatgacgatg acaagtgact agttctagag cggccgcgga tcccgcccct ctccctcccc
     3961 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     4021 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     4081 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     4141 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     4201 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     4261 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     4321 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     4381 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     4441 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     4501 ttgaaaaaca cgatgataag cttgccacaa cccacaagga gacgaccttc catgaccgag
     4561 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     4621 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     4681 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4741 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4801 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4861 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4921 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4981 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5041 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5101 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta gacgcgtctg
     5161 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5221 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5281 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5341 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5401 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5461 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5521 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     5581 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5641 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5701 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     5761 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct
     5821 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc
     5881 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     5941 ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt
     6001 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg
     6061 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca
     6121 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg
     6181 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca
     6241 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct
     6301 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg
     6361 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct
     6421 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     6481 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     6541 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     6601 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     6661 gaggccttga cattgctagc gtttaccgtc gacctctagc tagagcttgg cgtaatcatg
     6721 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6781 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6841 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6901 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6961 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7021 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7081 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7141 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7201 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7261 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7321 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7381 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7441 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7501 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7561 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7621 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7681 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7741 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7801 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7861 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7921 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7981 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8041 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8101 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8161 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8221 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8281 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8341 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8401 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8461 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8521 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8581 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8641 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8701 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8761 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8821 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8881 cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     8941 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     9001 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc
     9061 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct
     9121 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
     9181 ctacaaactt agtagttttt aaagaaattg tatttgttaa atatgtacta caaacttagt
     9241 agt
//
