LOCUS       V028321                 9580 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028321
VERSION     V028321
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..4473
                     /label="HSP90AB1(NM_007355)"
                     /note="HSP90AB1(NM_007355)"
                     /gene="HSP90AB1"
     CDS             4480..4506
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4550..4761
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4774..5042
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5069..5668
                     /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    5675..6263
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6337..6570
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6642..6763
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6782..6917
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6955..6971)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6979..6995
                     /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(7003..7033)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7048..7069
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7357..7945)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8116..8976)
                     /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(8977..9081)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9555..9571
                     /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 catgcctgag gaagtgcacc atggagagga ggaggtggag
     2341 acttttgcct ttcaggcaga aattgcccaa ctcatgtccc tcatcatcaa taccttctat
     2401 tccaacaagg agattttcct tcgggagttg atctctaatg cttctgatgc cttggacaag
     2461 attcgctatg agagcctgac agacccttcg aagttggaca gtggtaaaga gctgaaaatt
     2521 gacatcatcc ccaaccctca ggaacgtacc ctgactttgg tagacacagg cattggcatg
     2581 accaaagctg atctcataaa taatttggga accattgcca agtctggtac taaagcattc
     2641 atggaggctc ttcaggctgg tgcagacatc tccatgattg ggcagtttgg tgttggcttt
     2701 tattctgcct acttggtggc agagaaagtg gttgtgatca caaagcacaa cgatgatgaa
     2761 cagtatgctt gggagtcttc tgctggaggt tccttcactg tgcgtgctga ccatggtgag
     2821 cccattggca ggggtaccaa agtgatcctc catcttaaag aagatcagac agagtaccta
     2881 gaagagaggc gggtcaaaga agtagtgaag aagcattctc agttcatagg ctatcccatc
     2941 accctttatt tggagaagga acgagagaag gaaattagtg atgatgaggc agaggaagag
     3001 aaaggtgaga aagaagagga agataaagat gatgaagaaa aacccaagat cgaagatgtg
     3061 ggttcagatg aggaggatga cagcggtaag gataagaaga agaaaactaa gaagatcaaa
     3121 gagaaataca ttgatcagga agaactaaac aagaccaagc ctatttggac cagaaaccct
     3181 gatgacatca cccaagagga gtatggagaa ttctacaaga gcctcactaa tgactgggaa
     3241 gaccacttgg cagtcaagca cttttctgta gaaggtcagt tggaattcag ggcattgcta
     3301 tttattcctc gtcgggctcc ctttgacctt tttgagaaca agaagaaaaa gaacaacatc
     3361 aaactctatg tccgccgtgt gttcatcatg gacagctgtg atgagttgat accagagtat
     3421 ctcaatttta tccgtggtgt ggttgactct gaggatctgc ccctgaacat ctcccgagaa
     3481 atgctccagc agagcaaaat cttgaaagtc attcgcaaaa acattgttaa gaagtgcctt
     3541 gagctcttct ctgagctggc agaagacaag gagaattaca agaaattcta tgaggcattc
     3601 tctaaaaatc tcaagcttgg aatccacgaa gactccacta accgccgccg cctgtctgag
     3661 ctgctgcgct atcatacctc ccagtctgga gatgagatga catctctgtc agagtatgtt
     3721 tctcgcatga aggagacaca gaagtccatc tattacatca ctggtgagag caaagagcag
     3781 gtggccaact cagcttttgt ggagcgagtg cggaaacggg gcttcgaggt ggtatatatg
     3841 accgagccca ttgacgagta ctgtgtgcag cagctcaagg aatttgatgg gaagagcctg
     3901 gtctcagtta ccaaggaggg tctggagctg cctgaggatg aggaggagaa gaagaagatg
     3961 gaagagagca aggcaaagtt tgagaacctc tgcaagctca tgaaagaaat cttagataag
     4021 aaggttgaga aggtgacaat ctccaataga cttgtgtctt caccttgctg cattgtgacc
     4081 agcacctacg gctggacagc caatatggag cggatcatga aagcccaggc acttcgggac
     4141 aactccacca tgggctatat gatggccaaa aagcacctgg agatcaaccc tgaccacccc
     4201 attgtggaga cgctgcggca gaaggctgag gccgacaaga atgataaggc agttaaggac
     4261 ctggtggtgc tgctgtttga aaccgccctg ctatcttctg gcttttccct tgaggatccc
     4321 cagacccact ccaaccgcat ctatcgcatg atcaagctag gtctaggtat tgatgaagat
     4381 gaagtggcag cagaggaacc caatgctgca gttcctgatg agatcccccc tctcgagggc
     4441 gatgaggatg cgtctcgcat ggaagaagtc gatggatcct acccatacga cgtaccagat
     4501 tacgcttagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     4561 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     4621 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4681 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     4741 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     4801 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     4861 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     4921 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4981 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     5041 tccaagctgt gaccggcgcc tacgctagat gaccgagtac aagcccacgg tgcgcctcgc
     5101 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5161 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5221 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5281 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5341 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5401 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5461 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5521 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5581 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5641 gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg
     5701 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5761 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5821 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5881 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5941 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     6001 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6061 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     6121 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6181 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6241 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     6301 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6361 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6421 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6481 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6541 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6601 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6661 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6721 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     6781 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6841 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6901 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     6961 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7021 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7081 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7141 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7201 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7261 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7321 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7381 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7441 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7501 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7561 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7621 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7681 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7741 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7801 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7861 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7921 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7981 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8041 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8101 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8161 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8221 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8281 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8341 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8401 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8461 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8521 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8581 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8641 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8701 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8761 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8821 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8881 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8941 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     9001 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9061 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     9121 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     9181 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     9241 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     9301 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     9361 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     9421 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9481 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9541 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
