LOCUS       V036818                 8633 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036818
VERSION     V036818
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..3654
                     /label="TUBA1B(NM_006082)"
                     /note="TUBA1B(NM_006082)"
                     /gene="TUBA1B"
     CDS             3661..3687
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3691..3717
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3721..3747
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        3798..4009
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4022..4290
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4323..4721
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    4728..5316
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5390..5623
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5695..5816
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5835..5970
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6008..6024)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6032..6048
                     /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(6056..6086)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6101..6122
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6410..6998)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7169..8029)
                     /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(8030..8134)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8608..8624
                     /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 catgcgtgag tgcatctcca tccacgttgg ccaggctggt
     2341 gtccagattg gcaatgcctg ctgggagctc tactgcctgg aacacggcat ccagcccgat
     2401 ggccagatgc caagtgacaa gaccattggg ggaggagatg actccttcaa caccttcttc
     2461 agtgagacgg gcgctggcaa gcacgtgccc cgggctgtgt ttgtagactt ggaacccaca
     2521 gtcattgatg aagttcgcac tggcacctac cgccagctct tccaccctga gcagctcatc
     2581 acaggcaagg aagatgctgc caataactat gcccgagggc actacaccat tggcaaggag
     2641 atcattgacc ttgtgttgga ccgaattcgc aagctggctg accagtgcac cggtcttcag
     2701 ggcttcttgg ttttccacag ctttggtggg ggaactggtt ctgggttcac ctccctgctc
     2761 atggaacgtc tctcagttga ttatggcaag aagtccaagc tggagttctc catttaccca
     2821 gcaccccagg tttccacagc tgtagttgag ccctacaact ccatcctcac cacccacacc
     2881 accctggagc actctgattg tgccttcatg gtagacaatg aggccatcta tgacatctgt
     2941 cgtagaaacc tcgatatcga gcgcccaacc tacactaacc ttaaccgcct tattagccag
     3001 attgtgtcct ccatcactgc ttccctgaga tttgatggag ccctgaatgt tgacctgaca
     3061 gaattccaga ccaacctggt gccctacccc cgcatccact tccctctggc cacatatgcc
     3121 cctgtcatct ctgctgagaa agcctaccat gaacagcttt ctgtagcaga gatcaccaat
     3181 gcttgctttg agccagccaa ccagatggtg aaatgtgacc ctcgccatgg taaatacatg
     3241 gcttgctgcc tgttgtaccg tggtgacgtg gttcccaaag atgtcaatgc tgccattgcc
     3301 accatcaaaa ccaagcgcag catccagttt gtggattggt gccccactgg cttcaaggtt
     3361 ggcatcaact accagcctcc cactgtggtg cctggtggag acctggccaa ggtacagaga
     3421 gctgtgtgca tgctgagcaa caccacagcc attgctgagg cctgggctcg cctggaccac
     3481 aagtttgacc tgatgtatgc caagcgtgcc tttgttcact ggtacgtggg tgaggggatg
     3541 gaggaaggcg agttttcaga ggcccgtgaa gatatggctg cccttgagaa ggattatgag
     3601 gaggttggtg tggattctgt tgaaggagag ggtgaggaag aaggagagga atacctcgag
     3661 tacccatacg acgtacctga ttatgcaact tatccttatg atgtacctga ttatgctagc
     3721 tatccttatg atgtcccaga ctacgcatag ggatccgcgg ccgcgaagga tctgcgatcg
     3781 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     3841 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     3901 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     3961 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg
     4021 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg
     4081 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta
     4141 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc
     4201 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt
     4261 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctaggcca
     4321 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     4381 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     4441 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     4501 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     4561 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     4621 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     4681 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agtcgacaat caacctctgg
     4741 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     4801 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     4861 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     4921 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     4981 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5041 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5101 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     5161 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5221 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5281 agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact
     5341 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     5401 attcactccc aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac
     5461 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     5521 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     5581 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     5641 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     5701 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5761 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5821 gtctggctct agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5881 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5941 agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt
     6001 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6061 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6121 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6181 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6241 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6301 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6361 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6421 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6481 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6541 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6601 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6661 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6721 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6781 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6841 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6901 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6961 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7021 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7081 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7141 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7201 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7261 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7321 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7381 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7441 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7501 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7561 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7621 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7681 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7741 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7801 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7861 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7921 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7981 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8041 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8101 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8161 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     8221 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     8281 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     8341 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     8401 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     8461 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     8521 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     8581 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
