LOCUS       V028465                 8623 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028465
VERSION     V028465
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..3477
                     /label="BCAT2(NM_001190)"
                     /note="BCAT2(NM_001190)"
                     /gene="BCAT2"
     CDS             3484..3549
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3593..3804
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3817..4085
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4112..4711
                     /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    4718..5306
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5380..5613
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5685..5806
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5825..5960
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5998..6014)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6022..6038
                     /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(6046..6076)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6091..6112
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6400..6988)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7159..8019)
                     /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(8020..8124)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8598..8614
                     /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 catggccgca gccgctctgg ggcagatctg ggcacgaaag
     2341 cttctctctg tcccttggct tctgtgtggt cccagaagat atgcctcctc cagtttcaag
     2401 gctgcagacc tgcagctgga aatgacacag aagcctcata agaagcctgg ccccggcgag
     2461 cccctggtgt ttgggaagac atttaccgac cacatgctga tggtggaatg gaatgacaag
     2521 ggctggggcc agccccgaat ccagcccttc cagaacctca cgctgcaccc agcctcctcc
     2581 agcctccact actccctgca gctgtttgag ggcatgaagg cgttcaaagg caaagaccag
     2641 caggtgcgcc tcttccgccc ctggctcaac atggaccgga tgctgcgctc agccatgcgc
     2701 ctgtgcctgc cgagtttcga caagctggag ttgctggagt gcatccgccg gctcatcgaa
     2761 gtggacaagg actgggtccc cgatgccgcc ggcaccagcc tctatgtgcg gcctgtgctc
     2821 attgggaacg agccctcgct gggtgtcagc cagcccacgc gcgcgctcct gttcgtcatt
     2881 ctctgcccag tgggtgccta cttccctgga ggctccgtga ccccggtctc cctcctggcc
     2941 gacccagcct tcatccgggc ctgggtgggc ggggtcggca actacaagtt aggtgggaat
     3001 tatgggccca ccgtgttagt gcaacaggag gcactcaagc ggggctgtga acaggtcctc
     3061 tggctgtatg ggcccgacca ccagctcacc gaggtgggaa ccatgaacat ctttgtctac
     3121 tggacccacg aagatggggt gctggagctg gtgacgcccc cgctgaatgg tgttatcctg
     3181 cctggagtgg tcagacagag tctactggac atggctcaga cctggggtga gttccgggtg
     3241 gtggagcgca cgatcaccat gaagcagttg ctgcgggccc tggaggaggg ccgcgtgcgg
     3301 gaagtctttg gctcgggcac cgcttgccag gtctgcccag tgcaccgaat cctgtacaaa
     3361 gacaggaacc tccacattcc caccatggaa aatgggcctg agctgatcct ccgcttccag
     3421 aaggagctga aggagatcca gtacggaatc agagcccacg agtggatgtt cccggtggga
     3481 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3541 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     3601 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     3661 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     3721 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     3781 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     3841 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     3901 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     3961 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     4021 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     4081 agatccaagc tgtgaccggc gcctacgcta gatgaccgag tacaagccca cggtgcgcct
     4141 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     4201 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     4261 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4321 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4381 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4441 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4501 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4561 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4621 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4681 ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat caacctctgg attacaaaat
     4741 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4801 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4861 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4921 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4981 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5041 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5101 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     5161 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5221 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5281 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     5341 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     5401 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     5461 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     5521 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     5581 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     5641 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     5701 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5761 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     5821 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5881 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     5941 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6001 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6061 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6121 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6181 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6241 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6301 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6361 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6421 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6481 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6541 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6601 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6661 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6721 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6781 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6841 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6901 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6961 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7021 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7081 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7141 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7201 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7261 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7321 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7381 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7441 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7501 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7561 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7621 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7681 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7741 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7801 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7861 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7921 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7981 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8041 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8101 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8161 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8221 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8281 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     8341 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     8401 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     8461 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     8521 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     8581 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
