LOCUS       V035260                 7889 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035260
VERSION     V035260
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..2970
                     /label="UCHL1(NM_004181)"
                     /note="UCHL1(NM_004181)"
                     /gene="UCHL1"
     CDS             2977..3003
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        3054..3265
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3278..3546
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3579..3977
                     /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    3984..4572
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             4646..4879
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    4951..5072
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5091..5226
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5264..5280)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5288..5304
                     /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(5312..5342)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5357..5378
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5666..6254)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6425..7285)
                     /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(7286..7390)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     7864..7880
                     /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 catgcagctc aagccgatgg agatcaaccc cgagatgctg
     2341 aacaaagtgc tgtcccggct gggggtcgcc ggccagtggc gcttcgtgga cgtgctgggg
     2401 ctggaagagg agtctctggg ctcggtgcca gcgcctgcct gcgcgctgct gctgctgttt
     2461 cccctcacgg cccagcatga gaacttcagg aaaaagcaga ttgaagagct gaagggacaa
     2521 gaagttagtc ctaaagtgta cttcatgaag cagaccattg ggaattcctg tggcacaatc
     2581 ggacttattc acgcagtggc caataatcaa gacaaactgg gatttgagga tggatcagtt
     2641 ctgaaacagt ttctttctga aacagagaaa atgtcccctg aagacagagc aaaatgcttt
     2701 gaaaagaatg aggccataca ggcagcccat gatgccgtgg cacaggaagg ccaatgtcgg
     2761 gtagatgaca aggtgaattt ccattttatt ctgtttaaca acgtggatgg ccacctctat
     2821 gaacttgatg gacgaatgcc ttttccggtg aaccatggcg ccagttcaga ggacaccctg
     2881 ctgaaggacg ctgccaaggt ctgcagagaa ttcaccgagc gtgagcaagg agaagtccgc
     2941 ttctctgccg tggctctctg caaggcagcc ctcgagtacc catacgacgt accagattac
     3001 gcttgaggat ccgcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga
     3061 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
     3121 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     3181 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     3241 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     3301 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     3361 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     3421 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     3481 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     3541 cagatccaag ctgtgaccgg cgcctacgct aggccaccat ggccaagcct ttgtctcaag
     3601 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     3661 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     3721 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     3781 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     3841 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     3901 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     3961 atgtgtggga gggctaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     4021 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4081 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4141 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4201 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4261 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4321 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4381 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     4441 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4501 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4561 ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     4621 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat
     4681 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     4741 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     4801 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     4861 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     4921 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     4981 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5041 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     5101 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5161 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5221 gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg
     5281 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5341 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5401 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5461 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5521 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5581 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5641 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5701 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5761 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5821 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5881 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5941 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6001 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6061 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6121 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6181 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6241 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6301 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6361 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6421 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6481 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6541 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6601 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6661 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6721 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6781 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6841 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6901 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6961 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7021 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7081 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7141 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7201 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7261 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7321 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7381 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7441 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     7501 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     7561 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7621 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     7681 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     7741 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     7801 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     7861 gttgtaaaac gacggccagt gccaagctg
//
