LOCUS       V021858                 9287 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021858
VERSION     V021858
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             2826..3983
                     /label="BCAT1(NM_005504)"
                     /note="BCAT1(NM_005504)"
                     /gene="BCAT1"
     CDS             3990..4055
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4075..4574
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4595..5194
                     /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    5208..5796
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6003..6636
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6765..6781)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6789..6805
                     /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(6813..6843)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6858..6879
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7167..7755)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7926..8786)
                     /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(8787..8891)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8939..9073
                     /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 tcgctagcgc taccggactc agatctcgag
     2821 ccaccatgaa ggattgcagt aacggatgct ccgcagagtg taccggagaa ggaggatcaa
     2881 aagaggtggt ggggactttt aaggctaaag acctaatagt cacaccagct accattttaa
     2941 aggaaaaacc agaccccaat aatctggttt ttggaactgt gttcacggat catatgctga
     3001 cggtggagtg gtcctcagag tttggatggg agaaacctca tatcaagcct cttcagaacc
     3061 tgtcattgca ccctggctca tcagctttgc actatgcagt ggaattattt gaaggattga
     3121 aggcatttcg aggagtagat aataaaattc gactgtttca gccaaacctc aacatggata
     3181 gaatgtatcg ctctgctgtg agggcaactc tgccggtatt tgacaaagaa gagctcttag
     3241 agtgtattca acagcttgtg aaattggatc aagaatgggt cccatattca acatctgcta
     3301 gtctgtatat tcgtcctaca ttcattggaa ctgagccttc tcttggagtc aagaagccta
     3361 ccaaagccct gctctttgta ctcttgagcc cagtgggacc ttatttttca agtggaacct
     3421 ttaatccagt gtccctgtgg gccaatccca agtatgtaag agcctggaaa ggtggaactg
     3481 gggactgcaa gatgggaggg aattacggct catctctttt tgcccaatgt gaagcagtag
     3541 ataatgggtg tcagcaggtc ctgtggctct atggagagga ccatcagatc actgaagtgg
     3601 gaactatgaa tctttttctt tactggataa atgaagatgg agaagaagaa ctggcaactc
     3661 ctccactaga tggcatcatt cttccaggag tgacaaggcg gtgcattctg gacctggcac
     3721 atcagtgggg tgaatttaag gtgtcagaga gatacctcac catggatgac ttgacaacag
     3781 ccctggaggg gaacagagtg agagagatgt ttggctctgg tacagcctgt gttgtttgcc
     3841 cagtttctga tatactgtac aaaggcgaga caatacacat tccaactatg gagaatggtc
     3901 ctaagctggc aagccgcatc ttgagcaaat taactgatat ccagtatgga agagaagaga
     3961 gcgactggac aattgtgcta tccggatccg actacaaaga ccatgacggt gattataaag
     4021 atcatgacat cgactacaag gatgacgatg acaagtagtc tagataattc taccgggtag
     4081 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
     4141 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
     4201 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     4261 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     4321 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     4381 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     4441 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     4501 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     4561 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg
     4621 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     4681 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     4741 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4801 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4861 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4921 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4981 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5041 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5101 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5161 cacctggtgc atgacccgca agcccggtgc ctgaccgcgt ctggaacaat caacctctgg
     5221 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5281 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5341 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5401 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5461 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5521 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5581 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     5641 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5701 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5761 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga
     5821 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     5881 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     5941 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg
     6001 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     6061 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     6121 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     6181 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     6241 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     6301 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     6361 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     6421 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     6481 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6541 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6601 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6661 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct
     6721 agcgttttac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6781 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6841 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6901 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6961 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7021 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7081 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7141 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7201 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7261 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7321 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7381 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7441 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7501 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7561 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7621 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7681 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7741 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7801 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7861 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7921 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7981 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8041 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8101 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8161 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8221 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8281 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8341 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8401 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8461 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8521 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8581 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8641 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8701 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8761 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8821 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8881 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa
     8941 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     9001 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     9061 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     9121 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
     9181 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
     9241 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
