LOCUS       V045952                 9146 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045952
VERSION     V045952
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..4239
                     /label="HSPA8(NM_006597)"
                     /note="HSPA8(NM_006597)"
                     /gene="HSPA8"
     CDS             4249..4266
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        4311..4522
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4535..4803
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4836..5234
                     /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    5241..5829
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5903..6136
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6208..6329
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6348..6483
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6521..6537)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6545..6561
                     /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(6569..6599)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6614..6635
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6923..7511)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7682..8542)
                     /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(8543..8647)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9121..9137
                     /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 catgtccaag ggacctgcag ttggtattga tcttggcacc
     2341 acctactctt gtgtgggtgt tttccagcac ggaaaagtcg agataattgc caatgatcag
     2401 ggaaaccgaa ccactccaag ctatgtcgcc tttacggaca ctgaacggtt gatcggtgat
     2461 gccgcaaaga atcaagttgc aatgaacccc accaacacag tttttgatgc caaacgtctg
     2521 attggacgca gatttgatga tgctgttgtc cagtctgata tgaaacattg gccctttatg
     2581 gtggtgaatg atgctggcag gcccaaggtc caagtagaat acaagggaga gaccaaaagc
     2641 ttctatccag aggaggtgtc ttctatggtt ctgacaaaga tgaaggaaat tgcagaagcc
     2701 taccttggga agactgttac caatgctgtg gtcacagtgc cagcttactt taatgactct
     2761 cagcgtcagg ctaccaaaga tgctggaact attgctggtc tcaatgtact tagaattatt
     2821 aatgagccaa ctgctgctgc tattgcttac ggcttagaca aaaaggttgg agcagaaaga
     2881 aacgtgctca tctttgacct gggaggtggc acttttgatg tgtcaatcct cactattgag
     2941 gatggaatct ttgaggtcaa gtctacagct ggagacaccc acttgggtgg agaagatttt
     3001 gacaaccgaa tggtcaacca ttttattgct gagtttaagc gcaagcataa gaaggacatc
     3061 agtgagaaca agagagctgt aagacgcctc cgtactgctt gtgaacgtgc taagcgtacc
     3121 ctctcttcca gcacccaggc cagtattgag atcgattctc tctatgaagg aatcgacttc
     3181 tatacctcca ttacccgtgc ccgatttgaa gaactgaatg ctgacctgtt ccgtggcacc
     3241 ctggacccag tagagaaagc ccttcgagat gccaaactag acaagtcaca gattcatgat
     3301 attgtcctgg ttggtggttc tactcgtatc cccaagattc agaagcttct ccaagacttc
     3361 ttcaatggaa aagaactgaa taagagcatc aaccctgatg aagctgttgc ttatggtgca
     3421 gctgtccagg cagccatctt gtctggagac aagtctgaga atgttcaaga tttgctgctc
     3481 ttggatgtca ctcctctttc ccttggtatt gaaactgctg gtggagtcat gactgtcctc
     3541 atcaagcgta ataccaccat tcctaccaag cagacacaga ccttcactac ctattctgac
     3601 aaccagcctg gtgtgcttat tcaggtttat gaaggcgagc gtgccatgac aaaggataac
     3661 aacctgcttg gcaagtttga actcacaggc atacctcctg caccccgagg tgttcctcag
     3721 attgaagtca cttttgacat tgatgccaat ggtatactca atgtctctgc tgtggacaag
     3781 agtacgggaa aagagaacaa gattactatc actaatgaca agggccgttt gagcaaggaa
     3841 gacattgaac gtatggtcca ggaagctgag aagtacaaag ctgaagatga gaagcagagg
     3901 gacaaggtgt catccaagaa ttcacttgag tcctatgcct tcaacatgaa agcaactgtt
     3961 gaagatgaga aacttcaagg caagattaac gatgaggaca aacagaagat tctggacaag
     4021 tgtaatgaaa ttatcaactg gcttgataag aatcagactg ctgagaagga agaatttgaa
     4081 catcaacaga aagagctgga gaaagtttgc aaccccatca tcaccaagct gtaccagagt
     4141 gcaggaggca tgccaggagg aatgcctggg ggatttcctg gtggtggagc tcctccctct
     4201 ggtggtgctt cctcagggcc caccattgaa gaggttgatg cggccgcaca tcaccatcat
     4261 caccattgag cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg
     4321 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta
     4381 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     4441 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     4501 cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc
     4561 ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc
     4621 ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc
     4681 gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg
     4741 cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag
     4801 atccaagctg tgaccggcgc ctacgctagg ccaccatggc caagcctttg tctcaagaag
     4861 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     4921 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     4981 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     5041 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     5101 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     5161 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     5221 tgtgggaggg ctaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     5281 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     5341 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     5401 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     5461 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     5521 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     5581 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     5641 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     5701 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     5761 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     5821 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     5881 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg
     5941 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6001 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6061 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6121 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     6181 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     6241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6301 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     6361 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6421 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6481 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt
     6541 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6601 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7321 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8641 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     8701 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     8761 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     8821 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     8881 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     8941 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     9001 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     9061 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     9121 gtaaaacgac ggccagtgcc aagctg
//
