LOCUS       V046018                 9584 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046018
VERSION     V046018
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..3009
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             3040..4509
                     /label="CLK3(NM_003992)"
                     /note="CLK3(NM_003992)"
                     /gene="CLK3"
     promoter        4554..4765
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4778..5046
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5073..5672
                     /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    5679..6267
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6341..6574
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6646..6767
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6786..6921
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6959..6975)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6983..6999
                     /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(7007..7037)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7052..7073
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7361..7949)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8120..8980)
                     /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(8981..9085)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9559..9575
                     /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 aggataacat ggccatcatc
     2341 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     2401 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     2461 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     2521 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     2581 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     2641 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     2701 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     2761 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     2821 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     2881 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     2941 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     3001 ctgtacaagg gaggtggagg tagtggagga ggcggatcca tgcatcactg taagcgatac
     3061 cgctcccctg aaccagaccc gtacctgagc taccgatgga agaggaggag gtcctacagt
     3121 cgggaacatg aagggagact gcgatacccg tcccgaaggg agcctccccc acgaagatct
     3181 cggtccagaa gccatgaccg cctgccctac cagaggaggt accgggagcg ccgtgacagc
     3241 gatacatacc ggtgtgaaga gcggagccca tcctttggag aggactacta tggaccttca
     3301 cgttctcgtc atcgtcggcg atcgcgggag agggggccat accggacccg caagcatgcc
     3361 caccactgcc acaaacgccg caccaggtct tgtagcagcg cctcctcgag aagccaacag
     3421 agcagtaagc gcagcagccg gagtgtggaa gatgacaagg agggtcacct ggtgtgccgg
     3481 atcggcgatt ggctccaaga gcgatatgag attgtgggga acctgggtga aggcaccttt
     3541 ggcaaggtgg tggagtgctt ggaccatgcc agagggaagt ctcaggttgc cctgaagatc
     3601 atccgcaacg tgggcaagta ccgggaggct gcccggctag aaatcaacgt gctcaaaaaa
     3661 atcaaggaga aggacaaaga aaacaagttc ctgtgtgtct tgatgtctga ctggttcaac
     3721 ttccacggtc acatgtgcat cgcctttgag ctcctgggca agaacacctt tgagttcctg
     3781 aaggagaata acttccagcc ttacccccta ccacatgtcc ggcacatggc ctaccagctc
     3841 tgccacgccc ttagatttct gcatgagaat cagctgaccc atacagactt gaaaccagag
     3901 aacatcctgt ttgtgaattc tgagtttgaa accctctaca atgagcacaa gagctgtgag
     3961 gagaagtcag tgaagaacac cagcatccga gtggctgact ttggcagtgc cacatttgac
     4021 catgagcacc acaccaccat tgtggccacc cgtcactatc gcccgcctga ggtgatcctt
     4081 gagctgggct gggcacagcc ctgtgacgtc tggagcattg gctgcattct ctttgagtac
     4141 taccggggct tcacactctt ccagacccac gaaaaccgag agcacctggt gatgatggag
     4201 aagatcctag ggcccatccc atcacacatg atccaccgta ccaggaagca gaaatatttc
     4261 tacaaagggg gcctagtttg ggatgagaac agctctgacg gccggtatgt gaaggagaac
     4321 tgcaaacctc tgaagagtta catgctccaa gactccctgg agcacgtgca gctgtttgac
     4381 ctgatgagga ggatgttaga atttgaccct gcccagcgca tcacactggc cgaggccctg
     4441 ctgcacccct tctttgctgg cctgacccct gaggagcggt ccttccacac cagccgcaac
     4501 ccaagcagat gagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga
     4561 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
     4621 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     4681 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     4741 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     4801 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     4861 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     4921 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     4981 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     5041 cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc
     5101 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact
     5161 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc
     5221 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     5281 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     5341 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5401 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     5461 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5521 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5581 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5641 cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa
     5701 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     5761 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     5821 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     5881 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     5941 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     6001 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     6061 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     6121 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     6181 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     6241 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca
     6301 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     6361 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     6421 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6481 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6541 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     6601 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     6661 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6721 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     6781 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6841 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6901 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt
     6961 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7021 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7081 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7141 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7201 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7261 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7321 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7381 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7441 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7501 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7561 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7621 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7681 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7741 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7801 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7861 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7921 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7981 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     8041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8221 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8281 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8341 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8401 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8461 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8521 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8581 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8641 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8701 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8761 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8821 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8881 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8941 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9001 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     9061 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     9121 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     9181 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     9241 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     9301 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     9361 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     9421 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     9481 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     9541 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg
//
