LOCUS       V043572                 8852 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043572
VERSION     V043572
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..3018
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3055..3777
                     /label="CLIC1(NM_001287593)"
                     /note="CLIC1(NM_001287593)"
                     /gene="CLIC1"
     promoter        3822..4033
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4046..4314
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4341..4940
                     /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    4947..5535
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5609..5842
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5914..6035
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6054..6189
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6227..6243)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6251..6267
                     /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(6275..6305)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6320..6341
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6629..7217)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7388..8248)
                     /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(8249..8353)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8827..8843
                     /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 catggtgagc aagggcgagg agctgttcac cggggtggtg
     2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3001 atggacgagc tgtacaaggg cggcggaggg agtggaggag ggggaagtgg atccatggct
     3061 gaagaacaac cgcaggtcga attgttcgtg aaggctggca gtgatggggc caagattggg
     3121 aactgcccat tctcccagag actgttcatg gtactgtggc tcaagggagt caccttcaat
     3181 gttaccaccg ttgacaccaa aaggcggacc gagacagtgc agaagctgtg cccagggggg
     3241 cagctcccat tcctgctgta tggcactgaa gtgcacacag acaccaacaa gattgaggaa
     3301 tttctggagg cagtgctgtg ccctcccagg taccccaagc tggcagctct gaaccctgag
     3361 tccaacacag ctgggctgga catatttgcc aaattttctg cctacatcaa gaattcaaac
     3421 ccagcactca atgacaatct ggagaaggga ctcctgaaag ccctgaaggt tttagacaat
     3481 tacttaacat cccccctccc agaagaagtg gatgaaacca gtgctgaaga tgaaggtgtc
     3541 tctcagagga agtttttgga tggcaacgag ctcaccctgg ctgactgcaa cctgttgcca
     3601 aagttacaca tagtacaggt ggtgtgtaag aagtaccggg gattcaccat ccccgaggcc
     3661 ttccggggag tgcatcggta cttgagcaat gcctacgccc gggaagaatt cgcttccacc
     3721 tgtccagatg atgaggagat cgagctcgcc tatgagcaag tggcaaaggc cctcaaataa
     3781 gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc
     3841 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg
     3901 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     3961 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     4021 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc
     4081 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
     4141 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
     4201 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
     4261 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
     4321 gtgaccggcg cctacgctag atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     4381 acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     4441 gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     4501 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     4561 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     4621 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     4681 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     4741 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     4801 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     4861 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     4921 cccgcaagcc cggtgcctga gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     4981 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5041 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5101 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5161 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5221 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5281 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5341 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     5401 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5461 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5521 gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     5581 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa
     5641 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     5701 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     5761 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     5821 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     5881 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     5941 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6001 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     6061 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6121 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     6181 ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc
     6241 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6301 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6361 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6421 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6481 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6541 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6601 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6661 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6721 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6781 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6841 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6901 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6961 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7021 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7081 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7141 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7201 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7261 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7321 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7381 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7441 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7501 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7561 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7621 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7681 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7741 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7801 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7861 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7921 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7981 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8041 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8101 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8161 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8221 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8281 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8341 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     8401 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     8461 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     8521 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     8581 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     8641 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     8701 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     8761 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     8821 gacgttgtaa aacgacggcc agtgccaagc tg
//
