LOCUS       V028934                 8914 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028934
VERSION     V028934
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             2305..2331
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2332..3126
                     /label="Rspo1(NM_138683)"
                     /note="Rspo1(NM_138683)"
                     /gene="Rspo1"
     promoter        3884..4095
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4108..4376
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4403..5002
                     /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    5009..5597
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5671..5904
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5976..6097
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6116..6251
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6289..6305)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6313..6329
                     /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(6337..6367)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6382..6403
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6691..7279)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7450..8310)
                     /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(8311..8415)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8889..8905
                     /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 catgtaccca tacgacgtac cagattacgc tatgcggctt
     2341 gggctgtgcg tggtggccct ggttctgagc tggacacaca tcgccgtggg cagccggggg
     2401 atcaagggca agagacagag gcggatcagt gctgagggga gccaagcctg cgccaagggc
     2461 tgtgagctct gttcagaagt caacggttgc ctcaagtgct cgcccaagct cttcattctg
     2521 ctggagagga acgacatccg ccaggtgggc gtctgcctgc cgtcctgccc acctggatac
     2581 tttgatgccc gcaaccccga catgaacaaa tgcatcaaat gcaagatcga gcactgtgag
     2641 gcctgcttca gccacaactt ctgcaccaag tgtcaggagg gcttgtactt acacaagggc
     2701 cgctgctatc cagcctgccc tgagggctct acagccgcta acagcaccat ggagtgcggc
     2761 agtcctgcac aatgtgaaat gagcgagtgg tccccgtggg gaccctgctc caagaagagg
     2821 aagctgtgcg gtttccggaa gggatcggaa gagcggacac gcagagtgct ccatgctccc
     2881 gggggagacc acaccacctg ctccgacacc aaagagaccc gcaagtgtac cgtgcgcagg
     2941 acgccctgcc cagaggggca gaagaggagg aaggggggcc agggccggag ggagaatgcc
     3001 aacaggcatc cggccaggaa gaacagcaag gagccgggct ccaactctcg gagacacaaa
     3061 gggcaacagc agccacagcc agggacaaca gggccactca catcagtagg acctacctgg
     3121 gcacagggtg gaggtggatc agagcccagg ggccccacca tcaagccctg ccccccctgc
     3181 aagtgccccg cccccaacct gctgggcggc cccagcgtgt tcatcttccc ccccaagatc
     3241 aaggacgtgc tgatgatcag cctgagcccc atcgtgacct gcgtggtggt ggacgtgagc
     3301 gaggacgacc ccgacgtgca gatcagctgg ttcgtgaaca acgtggaggt gcacaccgcc
     3361 cagacccaga cccacaggga ggactacaac agcaccctga gggtggtgag cgccctgccc
     3421 atccagcacc aggactggat gagcggcaag gagttcaagt gcaaggtgaa caacaaggac
     3481 ctgcccgccc ccatcgagag gaccatcagc aagcccaagg gcagcgtgag ggccccccag
     3541 gtgtacgtgc tgcccccccc cgaggaggag atgaccaaga agcaggtgac cctgacctgc
     3601 atggtgaccg acttcatgcc cgaggacatc tacgtggagt ggaccaacaa cggcaagacc
     3661 gagctgaact acaagaacac cgagcccgtg ctggacagcg acggcagcta cttcatgtac
     3721 agcaagctga gggtggagaa gaagaactgg gtggagagga acagctacag ctgcagcgtg
     3781 gtgcacgagg gcctgcacaa ccaccacacc accaagagct tcagcaggac ccccggcaag
     3841 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     3901 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     3961 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     4021 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     4081 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     4141 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     4201 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     4261 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     4321 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag
     4381 ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4441 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4501 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4561 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4621 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4681 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4741 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4801 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4861 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4921 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4981 gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     5041 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5101 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5161 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5221 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5281 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5341 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5401 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5461 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5521 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5581 ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5641 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     5701 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5761 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     5821 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     5881 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     5941 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6001 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6061 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6121 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6181 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6241 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     6301 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6361 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6421 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6481 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7261 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7321 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7381 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7441 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7501 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7561 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7621 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7681 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7741 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7801 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7861 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7921 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7981 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8041 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8101 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8161 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8221 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8281 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8341 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8401 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8461 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8521 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     8581 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     8641 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     8701 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     8761 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     8821 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     8881 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
