LOCUS       V037394                 9952 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037394
VERSION     V037394
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"
     regulatory      2296..2305
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2305..2394
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             2401..4863
                     /label="EVI5(NM_001308248)"
                     /note="EVI5(NM_001308248)"
                     /gene="EVI5"
     promoter        4922..5133
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5146..5414
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5441..6040
                     /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    6047..6635
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6709..6942
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7014..7135
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7154..7289
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7327..7343)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7351..7367
                     /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(7375..7405)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7420..7441
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7729..8317)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8488..9348)
                     /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(9349..9453)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9927..9943
                     /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 catggaacaa aaactaatat cagaggaaga tttggaacag
     2341 aaactaatca gtgaagaaga tttagagcaa aagttaattt ccgaggagga cttactcgag
     2401 atggttacca acaaaatgac tgctgccttt agaaacccta gtgggaaaca ggtggcgaca
     2461 gacaaagttg cagaaaagct gagctctact ctctcatggg tgaagaacac agtatcgcat
     2521 acagtcagtc agatggccag tcaggtggca agtccatcta cttcattaca taccacatcc
     2581 tcatctacca cactatcaac accagccctt tcaccatctt ccccatcaca gttgagtcca
     2641 gacgacttag aactcctggc taaactggaa gaacagaata gattgttaga aacggatagt
     2701 aagtctttaa gatctgtaaa tgggtcaaga agaaacagtg gctcttctct tgtgtcgagt
     2761 tcatcagcct ctagcaacct cagtcacctt gaagaagatt cttggattct ttggggaaga
     2821 attgttaatg aatgggaaga tgtacgcaaa aagaaggaaa agcaagttaa ggaacttgtt
     2881 cataaaggga taccccatca ctttagagca atagtttggc aacttttatg cagtgcacaa
     2941 agtatgccaa ttaaggatca gtattcagaa ctcctgaaaa tgacctcgcc ttgtgaaaaa
     3001 ttgatccgaa gggacattgc tagaacttac cctgaacaca acttttttaa ggaaaaagat
     3061 agccttggac aggaggtttt atttaatgta atgaaggctt actctttagt agatcgtgag
     3121 gttggttact gtcaaggaag tgcttttata gttggattgt tgcttatgca gatgccagaa
     3181 gaagaagctt tctgtgtatt tgttaaatta atgcaagatt atagacttcg tgaacttttt
     3241 aaaccaagta tggcagaatt gggcctttgt atgtaccagt ttgaatgtat gatacaggag
     3301 catcttccag agctctttgt acattttcaa tctcagagtt ttcatacctc aatgtatgca
     3361 tcatcctggt ttctgactat ctttcttaca acttttccac taccaattgc aacaaggata
     3421 tttgatatct ttatgtctga gggtttagaa atagtgtttc gtgtaggatt agcacttctt
     3481 cagatgaatc aggcagaact gatgcaactt gacatggaag ggatgttaca gcactttcaa
     3541 aaggtcattc cacatcagtt tgatggtgtc ccagacaagc taatccaagc agcttaccaa
     3601 gtcaaataca attcaaaaaa aatgaaaaag cttgaaaagg aatacactac aataaaaacg
     3661 aaagaaatgg aagagcaagt tgaaattaaa aggttacgca cagaaaatag acttttaaaa
     3721 cagcgcatcg agacattaga aaaagaaagt gcttccttgg cagatagatt gatacagcat
     3781 aaatgcagtt ccaactacaa cgaagatttt gtgctacagc tagagaagga attggtccaa
     3841 gcccgactga gtgaagctga gtctcagtgt gcattaaaag agatgcagga taaagtcttg
     3901 gatatagaga agaggaataa ctcccttcct gatgagaata atattgcaag gcttcaggaa
     3961 gaactcattg ctgtgaaact tagagaagca gaagccatta tgggtttgaa agaacttaga
     4021 cagcaagtca aggatttaga ggaacactgg cagcgccact tagctcgtac tactgggaga
     4081 tggaaagacc cacccaagaa aaatgctatg aatgagttac aagatgaact gatgaccatt
     4141 cgacttagag aagctgaaac acaagcagaa ataagagaaa taaaacaaag gatgatggaa
     4201 atggaaacac agaaccagat caatagtaac catcttcgaa gagcagaaca agaggtgatt
     4261 agcctacagg agaaagtgca gtatctttct gcacagaaca aaggactcct tactcaatta
     4321 agtgaagcaa agcgtaaaca agcagagatt gaatgcaaga ataaggaaga agtgatggct
     4381 gtgaggcttc gggaagcaga tagcatagct gctgtggctg aactacgaca acacattgct
     4441 gagcttgaaa tccagaaaga agaaggaaag cttcaaggac agcttaacaa gtctgattct
     4501 aaccagtata ttggggaact gaaagatcag atagcagagc tgaatcatga gctcaggtgc
     4561 ctaaaaggcc agaggggctt ctcaggccaa cctccttttg atggaatcca cattgtcaac
     4621 catttaatag gagatgatga atcattccat tcctccgatg aagattttat agataattcc
     4681 ttacaggaaa ctggtgttgg ttttcctttg cacggaaaat ctggttcgat gtctttggac
     4741 cccgcagtgg cagatggtag tgagagcgaa acagaagaca gtgtgctgga gaccagagag
     4801 agcaaccaag tggttcaaaa ggagcggccc ccgagaagaa gagagtcgta ttcaaccact
     4861 gtctgaggat ccgatatcta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     4921 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     4981 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     5041 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     5101 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     5161 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     5221 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     5281 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     5341 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     5401 cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac
     5461 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     5521 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     5581 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5641 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5701 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5761 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5821 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5881 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5941 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     6001 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga
     6061 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     6121 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     6181 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     6241 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     6301 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     6361 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     6421 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     6481 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     6541 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     6601 gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt
     6661 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     6721 ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     6781 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6841 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6901 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6961 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     7021 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     7081 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     7141 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     7201 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     7261 gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta
     7321 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7381 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7441 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7501 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7561 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7621 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7681 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7741 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7801 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7861 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7921 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7981 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8041 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8101 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8161 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8221 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8281 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8341 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8401 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8461 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8521 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8581 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8641 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8701 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8761 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8821 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8881 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8941 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     9001 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9061 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9121 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9181 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9241 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9301 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9361 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9421 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9481 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     9541 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     9601 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     9661 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     9721 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     9781 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     9841 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     9901 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
