LOCUS       V029515                 9804 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029515
VERSION     V029515
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        1960..2171
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             2184..2452
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             2486..4675
                     /label="DTL(NM_016448)"
                     /note="DTL(NM_016448)"
                     /gene="DTL"
     CDS             4682..4747
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4773..5272
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5293..5892
                     /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    5899..6487
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6561..6794
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6866..6987
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7006..7141
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7179..7195)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7203..7219
                     /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(7227..7257)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7272..7293
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7581..8169)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8340..9200)
                     /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(9201..9305)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9779..9795
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca
     1981 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
     2041 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     2101 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     2161 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
     2221 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
     2281 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
     2341 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
     2401 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
     2461 gaccggcgcc tactctagag ccaccatgct cttcaattcg gtgctccgcc agccccagct
     2521 tggcgtcctg agaaatggat ggtcttcaca ataccctctt caatcccttc tgactggtta
     2581 tcagtgcagt ggtaatgatg aacacacttc ttatggagaa acaggagtcc cagttcctcc
     2641 ttttggatgt accttctctt ctgctcccaa tatggaacat gtactagcag ttgccaatga
     2701 agaaggcttt gttcgattgt ataacacaga atcacaaagt ttcagaaaga agtgcttcaa
     2761 agaatggatg gctcactgga atgccgtctt tgacctggcc tgggttcctg gtgaacttaa
     2821 acttgttaca gcagcaggtg atcaaacagc caaattttgg gacgtaaaag ctggtgagct
     2881 gattggaaca tgcaaaggtc atcaatgcag cctcaagtca gttgcctttt ctaagtttga
     2941 gaaagctgta ttctgtacgg gtggaagaga tggcaacatt atggtctggg ataccaggtg
     3001 caacaaaaaa gatgggtttt ataggcaagt gaatcaaatc agtggagctc acaatacctc
     3061 agacaagcaa accccttcaa aacccaagaa gaaacagaat tcaaaaggac ttgctccttc
     3121 tgtggatttc cagcaaagtg ttactgtggt cctctttcaa gacgagaata ccttagtctc
     3181 agcaggagct gtggatggga taatcaaagt atgggattta cgtaagaatt atactgctta
     3241 tcgacaagaa cccatagcat ccaagtcttt cctgtaccca ggtagcagca ctcgaaaact
     3301 tggatattca agtctgattt tggattccac tggctctact ttatttgcta attgcacaga
     3361 cgataacatc tacatgttta atatgactgg gttgaagact tctccagtgg ctattttcaa
     3421 tggacaccag aactctacct tttatgtaaa atccagcctt agtccagatg accagttttt
     3481 agtcagtggc tcaagtgatg aagctgccta catatggaag gtctccacac cctggcaacc
     3541 tcctactgtg ctcctgggtc attctcaaga ggtcacgtct gtgtgctggt gtccatctga
     3601 cttcacaaag attgctacct gttctgatga caatacacta aaaatctggc gcttgaatag
     3661 aggcttagag gagaaaccag gaggtgataa actttccacg gtgggttggg cctctcagaa
     3721 gaaaaaagag tcaagacctg gcctagtaac agtaacgagt agccagagta ctcctgccaa
     3781 agcccccagg gcaaagtgca atccatccaa ttcttccccg tcatccgcag cttgtgcccc
     3841 aagctgtgct ggagacctcc ctcttccttc aaatactcct acgttctcta ttaaaacctc
     3901 tcctgccaag gcccggtctc ccatcaacag aagaggctct gtctcctccg tctctcccaa
     3961 gccaccttca tctttcaaga tgtcgattag aaactgggtg acccgaacac cttcctcatc
     4021 accacccatc actccacctg cttcggagac caagatcatg tctccgagaa aagcccttat
     4081 tcctgtgagc cagaagtcat cccaagcaga ggcttgctct gagtctagaa atagagtaaa
     4141 gaggaggcta gactcaagct gtctggagag tgtgaaacaa aagtgtgtga agagttgtaa
     4201 ctgtgtgact gagcttgatg gccaagttga aaatcttcat ttggatctgt gctgccttgc
     4261 tggtaaccag gaagacctta gtaaggactc tctaggtcct accaaatcaa gcaaaattga
     4321 aggagctggt accagtatct cagagcctcc gtctcctatc agtccgtatg cttcagaaag
     4381 ctgtggaacg ctacctcttc ctttgagacc ttgtggagaa gggtctgaaa tggtaggcaa
     4441 agagaatagt tccccagaga ataaaaactg gttgttggcc atggcagcca aacggaaggc
     4501 tgagaatcca tctccacgaa gtccgtcatc ccagacaccc aattccagga gacagagcgg
     4561 aaagaaattg ccaagcccgg tcaccatcac gcccagctcc atgaggaaaa tctgcacata
     4621 cttccataga aagtcccagg aggacttctg tggtcctgaa cactcaacag aattagctag
     4681 cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     4741 tgacaagtga attcgaagga tccgcggccg ctgggtaggg gaggcgcttt tcccaaggca
     4801 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     4861 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     4921 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     4981 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     5041 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     5101 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
     5161 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     5221 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
     5281 cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     5341 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     5401 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     5461 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     5521 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     5581 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5641 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     5701 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     5761 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     5821 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     5881 cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5941 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6001 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6061 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6121 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6181 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6241 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     6301 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6361 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6421 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     6481 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6541 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct
     6601 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6661 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6721 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6781 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     6841 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     6901 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6961 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     7021 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     7081 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     7141 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga
     7201 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     7261 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     7321 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     7381 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     7441 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     7501 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7561 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7621 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7681 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7741 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7801 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7861 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7921 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7981 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     8041 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8101 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8161 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8221 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8281 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8341 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8401 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8461 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8521 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8581 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8641 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8701 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8761 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8821 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8881 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8941 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9001 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9061 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9121 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9181 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9241 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9301 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     9361 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     9421 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     9481 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     9541 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     9601 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     9661 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     9721 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     9781 aaaacgacgg ccagtgccaa gctg
//
