LOCUS       V031546                 9704 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031546
VERSION     V031546
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             2290..3846
                     /label="STK3(NM_001256312)"
                     /note="STK3(NM_001256312)"
                     /gene="STK3"
     CDS             3883..4599
                     /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             4606..4623
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        4674..4885
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4898..5166
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5193..5792
                     /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    5799..6387
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6461..6694
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6766..6887
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6906..7041
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7079..7095)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7103..7119
                     /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(7127..7157)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7172..7193
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7481..8069)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8240..9100)
                     /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(9101..9205)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9679..9695
                     /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 agagccacca tgttacaact catggacagt ggcattacca tctgccttag gaacggagca
     2341 gcctctgtct tcaagaaaaa agaatggagc acacaaggag aagaaaacaa gcaagacagt
     2401 aaactaaaaa agctgagtga agacagtttg actaagcagc ctgaagaagt ttttgatgta
     2461 ttagagaagc ttggagaagg gtcttatgga agtgtattta aagcaataca caaggaatcc
     2521 ggtcaagttg tcgcaattaa acaagtacct gttgaatcag atcttcagga aataatcaaa
     2581 gaaatttcca taatgcagca atgtgacagc ccatatgttg taaagtacta tggcagttat
     2641 tttaagaata cagacctctg gattgttatg gagtactgtg gcgctggctc tgtctcagac
     2701 ataattagat tacgaaacaa gacattaata gaagatgaaa ttgcaaccat tcttaaatct
     2761 acattgaaag gactagaata tttgcacttt atgagaaaaa tacacagaga tataaaagct
     2821 ggaaatattc tcctcaatac agaaggacat gcaaaattgg cagattttgg agtggctggt
     2881 cagttaacag atacaatggc aaaacgcaat actgtaatag gaactccatt ttggatggct
     2941 cctgaggtga ttcaagaaat aggctataac tgtgtggccg acatctggtc ccttggcatt
     3001 acttctatag aaatggctga aggaaaacct ccttatgctg atatacatcc aatgagggct
     3061 atttttatga ttcccacaaa tccaccacca acattcagaa agccagaact ttggtccgat
     3121 gatttcaccg attttgttaa aaagtgtttg gtgaagaatc ctgagcagag agctactgca
     3181 acacaacttt tacagcatcc ttttatcaag aatgccaaac ctgtatcaat attaagagac
     3241 ctgatcacag aagctatgga gatcaaagct aaaagacatg aggaacagca acgagaattg
     3301 gaagaggaag aagaaaattc ggatgaagat gagctggatt cccacaccat ggtgaagact
     3361 agtgtggaga gtgtgggcac catgcgggcc acaagcacga tgagtgaagg ggcccagacc
     3421 atgattgaac ataatagcac gatgttggaa tccgacttgg ggaccatggt gataaacagt
     3481 gaggatgagg aagaagaaga tggaactatg aaaagaaatg caacctcacc acaagtacaa
     3541 agaccatctt tcatggacta ctttgataag caagacttca agaataagag tcacgaaaac
     3601 tgtaatcaga acatgcatga acccttccct atgtccaaaa acgtttttcc tgataactgg
     3661 aaagttcctc aagatggaga ctttgacttt ttgaaaaatc taagtttaga agaactacag
     3721 atgcggttaa aagcactgga ccccatgatg gaacgggaga tagaagaact tcgtcagaga
     3781 tacactgcga aaagacagcc cattctggat gcgatggatg caaagaaaag aaggcagcaa
     3841 aactttgaat tcggaggtgg cggaagcggc ggtggaggta gtatggtgag caagggcgag
     3901 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3961 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     4021 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     4081 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     4141 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     4201 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4261 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4321 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     4381 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4441 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4501 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4561 gccgccggga tcactctcgg catggacgag ctgtacaagc tcgagcacca ccaccaccac
     4621 cactgaggat ccgcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga
     4681 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
     4741 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     4801 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     4861 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     4921 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     4981 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     5041 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     5101 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     5161 cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc
     5221 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact
     5281 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc
     5341 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     5401 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     5461 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5521 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     5581 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5641 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5701 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5761 cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa
     5821 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     5881 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     5941 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     6001 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     6061 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     6121 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     6181 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     6241 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     6301 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     6361 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca
     6421 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     6481 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     6541 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6601 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6661 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     6721 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     6781 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6841 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     6901 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6961 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     7021 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt
     7081 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7141 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7201 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7261 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7321 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7381 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7921 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7981 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     8041 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     8101 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     8161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8281 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8341 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8401 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8461 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8521 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8581 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8641 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8701 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8761 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8821 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8881 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8941 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     9001 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     9061 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9121 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     9181 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     9241 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     9301 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     9361 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     9421 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     9481 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     9541 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     9601 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     9661 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg
//
