LOCUS       V034281                 9880 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034281
VERSION     V034281
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..4734
                     /label="Postn(NM_015784)"
                     /note="Postn(NM_015784)"
                     /gene="Postn"
     CDS             4741..4806
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4850..5061
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5074..5342
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5369..5968
                     /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    5975..6563
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6637..6870
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6942..7063
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7082..7217
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7255..7271)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7279..7295
                     /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(7303..7333)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7348..7369
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7657..8245)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8416..9276)
                     /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(9277..9381)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9855..9871
                     /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 catggttcct ctcctgccct tatatgctct gctgctgctg
     2341 ttcctgtgtg atattaaccc tgcaaatgcc aacagttact atgacaaggt cctggctcac
     2401 agccgcatca ggggtcggga tcagggccca aacgtctgtg ccctccagca aattctgggc
     2461 accaaaaaga aatacttcag ctcctgtaag aactggtatc aaggtgctat ctgcgggaag
     2521 aaaaccactg tgctatatga atgctgccct ggctatatga gaatggaagg gatgaaaggc
     2581 tgccccgcag tgatgcctat tgaccatgtt tatggcacgc tgggcattgt gggagccact
     2641 accactcagc actactccga tgtctcgaag ctgagagaag agattgaagg aaaagggtca
     2701 tacacgtact tcgcgccgag taacgaggct tgggagaacc tggattctga cattcgcaga
     2761 ggactggaga acaatgtcaa tgttgagcta ctgaatgcct tacacagcca catggttaat
     2821 aagagaatgt taaccaagga cctgaaacac ggcatggtta ttccttcaat gtacaacaat
     2881 ctggggcttt ttattaacca ttatcccaat ggggttgtca ctgtgaactg tgctcgagtc
     2941 atccatggga accagattgc cacaaatggt gtcgtccatg tcattgaccg tgtcctgaca
     3001 caaattggta cctccatcca agacttcctt gaagcagaag acgacctttc atcatttaga
     3061 gcagccgcca tcacctctga cctcttggag tcccttggaa gagatggtca cttcacgctc
     3121 tttgctccca ccaatgaagc tttcgagaaa ctgccacgag gtgtcctaga aaggatcatg
     3181 ggagacaaag tggcttctga agctctcatg aagtaccaca tcctaaatac cctccagtgc
     3241 tctgaggcca tcactggagg agccgtgttt gagaccatgg aaggaaacac tattgagata
     3301 gggtgcgaag gggacagtat ctccattaac ggaatcaaga tggtgaacaa gaaagacatt
     3361 gtgactaaga atggtgtcat ccacctgatt gatgaagtcc tcattcctga ttctgccaaa
     3421 caagttattg agctggctgg aaaacagcaa accactttca ccgacctggt agcccaatta
     3481 ggcttggcat cctctctgaa gccagatgga gagtacacct tattagcacc tgtgaacaat
     3541 gcgttctctg atgacactct gagcatggac caacgccttc ttaagctaat tctgcaaaat
     3601 cacatattga aagtaaaagt tggccttagc gacctctaca atggacagat actggaaacc
     3661 attggaggca aacaactccg agtctttgtg tatcggacgg ctatctgcat agaaaactca
     3721 tgcatggtga gaggaagcaa gcagggaagg aatggtgcca ttcacatatt ccgagaaatc
     3781 atccaaccag cagagaaatc cctgcacgac aagctgcggc aagacaagcg ctttagcatc
     3841 ttcctcagcc tccttgaagc tgcagatttg aaagatctcc tgacacagcc cggagattgg
     3901 accttgtttg caccaaccaa tgatgccttc aagggaatga ctagcgaaga aagggagctt
     3961 ctgattgggg ataaaaatgc tctccaaaac atcattcttt atcacctgac cccaggggtt
     4021 tatattggaa agggattcga acccggagtc actaatatcc tgaagaccac acagggaagc
     4081 aaaatctatc tgaaaggagt aaacgaaacg cttctagtga atgagttgaa gtccaaagaa
     4141 tctgacatca tgacgacaaa tggtgtcatc cacgtcgtgg acaaactcct ctatccagca
     4201 gatattccag ttggaaatga tcagctcttg gaattactga acaaactgat aaaatacatc
     4261 caaatcaagt ttgttcgtgg cagcaccttc aaagaaatcc ccatgactgt ctatagacct
     4321 gcaatgacga agatccaaat tgaaggtgat cccgacttca ggctgattaa agaaggcgaa
     4381 acggtgacag aagtgatcca cggagagcca gtcattaaaa agtacaccaa aatcatagat
     4441 ggagttcctg ttgaaataac tgaaaaacag actcgggaag aacgaatcat tacaggtcct
     4501 gagataaaat ataccaggat ttccacagga ggtggagaaa caggagagac cttgcagaaa
     4561 ttcttgcaaa aagaggtctc caaggtcaca aagttcattg aaggtggcga tggtcactta
     4621 tttgaagatg aggagattaa aagactgctt cagggagaca cacctgcaaa gaagatacca
     4681 gccaacaaaa gggttcaagg gcctagaaga cgatcaagag aaggccgttc tcagggatcc
     4741 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     4801 gacaagtagc ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     4861 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     4921 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     4981 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     5041 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     5101 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     5161 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     5221 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     5281 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     5341 tccaagctgt gaccggcgcc tacgctagat gaccgagtac aagcccacgg tgcgcctcgc
     5401 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5461 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5521 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5581 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5641 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5701 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5761 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5821 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5881 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5941 gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg
     6001 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     6061 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     6121 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     6181 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     6241 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     6301 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6361 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     6421 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6481 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6541 ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg
     6601 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6661 gaaaataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6721 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6781 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6841 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6901 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     7021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     7081 tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     7141 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     7201 aggctttttt ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata
     7261 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7321 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7381 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7441 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7501 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7561 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7621 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7681 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7741 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7801 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7861 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7921 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7981 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8041 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     8101 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     8161 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     8221 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8281 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8341 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8401 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8461 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8521 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8581 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8641 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8701 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8761 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8821 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8881 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8941 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9001 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     9061 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     9121 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     9181 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9241 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     9301 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9361 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     9421 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     9481 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     9541 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     9601 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     9661 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     9721 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9781 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9841 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
