LOCUS       V035271                 9244 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035271
VERSION     V035271
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..4098
                     /label="Esr1(NM_007956)"
                     /note="Esr1(NM_007956)"
                     /gene="Esr1"
     CDS             4105..4170
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4214..4425
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4438..4706
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4733..5332
                     /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    5339..5927
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6001..6234
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6306..6427
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6446..6581
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6619..6635)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6643..6659
                     /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(6667..6697)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6712..6733
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7021..7609)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7780..8640)
                     /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(8641..8745)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9219..9235
                     /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 catgaccatg acccttcaca ccaaagcctc gggaatggcc
     2341 ttgctgcacc agatccaagg gaacgagctg gagcccctca accgcccgca gctcaagatg
     2401 cccatggaga gggccctggg cgaggtatac gtggacaaca gcaagcccac tgtgttcaac
     2461 taccccgagg gcgccgccta cgagttcaac gccgccgccg ccgccgccgc cgccgcctcg
     2521 gcgccggtct acggccagtc gggcatcgcc tacggccccg ggtcggaggc ggccgccttc
     2581 agtgccaaca gcctgggggc tttcccccag ctcaacagcg tgtcgcctag cccgctgatg
     2641 ctgctgcacc cgccgccgca gctgtctcct ttcctgcacc cgcacggcca gcaggtgccc
     2701 tactacctgg agaacgagcc cagcgcctac gccgtgcgcg acaccggccc tcccgccttc
     2761 tacaggtcta attctgacaa tcgacgccag aatggccgag agagactgtc cagcagtaac
     2821 gagaaaggaa acatgatcat ggagtctgcc aaggagactc gctactgtgc cgtgtgcaat
     2881 gactatgcct ctggctacca ttatggggtc tggtcctgcg aaggctgcaa ggctttcttt
     2941 aagagaagca ttcaaggaca caatgactac atgtgtccag ctacaaacca atgcaccatt
     3001 gacaagaacc ggaggaagag ttgccaggcc tgtcggctgc gcaagtgtta cgaagtgggc
     3061 atgatgaaag gcggcatacg gaaagaccgc cgaggaggga gaatgttgaa gcacaagcgt
     3121 cagagagatg acttggaagg ccgaaatgaa atgggtgctt caggagacat gagggctgcc
     3181 aacctttggc caagccctct tgtgattaag cacactaaga agaatagccc tgccttgtcc
     3241 ttgacagctg accagatggt cagtgccttg ttggatgctg aaccgcccat gatctattct
     3301 gaatatgatc cttctagacc cttcagtgaa gcctcaatga tgggcttatt gaccaaccta
     3361 gcagataggg agctggttca tatgatcaac tgggcaaaga gagtgccagg ctttggggac
     3421 ttgaatctcc atgatcaggt ccaccttctc gagtgtgcct ggctggagat tctgatgatt
     3481 ggtctcgtct ggcgctccat ggaacacccg gggaagctcc tgtttgctcc taacttgctc
     3541 ctggacagga atcaaggtaa atgtgtggaa ggcatggtgg agatctttga catgttgctg
     3601 gctacgtcaa gtcggttccg catgatgaac ctgcagggag aagagtttgt gtgcctcaaa
     3661 tccatcattt tgcttaattc cggagtgtac acgtttctgt ccagcacctt gaagtctctg
     3721 gaagagaagg accacatcca ccgtgtcctg gacaagatca cagacacttt gatccacctg
     3781 atggccaaag ctggcctgac tctgcagcag cagcatcgcc gcctagctca gctccttctc
     3841 attctttccc atatccggca catgagtaac aaaggcatgg agcatctcta caacatgaaa
     3901 tgcaagaacg ttgtgcccct ctatgacctg ctcctggaga tgttggatgc ccaccgcctt
     3961 catgccccag ccagtcgcat gggagtgccc ccagaggagc ccagccagac ccagctggcc
     4021 accaccagct ccacttcagc acattcctta caaacctact acataccccc ggaagcagag
     4081 ggcttcccca acacgatcgg atccgactac aaagaccatg acggtgatta taaagatcat
     4141 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc
     4201 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     4261 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     4321 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     4381 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     4441 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     4501 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     4561 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     4621 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     4681 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga
     4741 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     4801 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     4861 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4921 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4981 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5041 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5101 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5161 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5221 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5281 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa
     5341 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     5401 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     5461 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     5521 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     5581 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     5641 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5701 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     5761 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5821 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5881 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa
     5941 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     6001 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc
     6061 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     6121 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     6181 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     6241 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6301 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6361 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6421 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc
     6481 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6541 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg
     6601 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6661 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6721 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6781 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7441 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7681 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8761 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8821 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     8881 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     8941 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     9001 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     9061 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     9121 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     9181 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     9241 gctg
//
