LOCUS       V024631                 9926 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024631
VERSION     V024631
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     regulatory      2279..2288
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2288..2353
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2366..4660
                     /label="Cd248(NM_054042)"
                     /note="Cd248(NM_054042)"
                     /gene="Cd248"
     promoter        4729..4940
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4953..5221
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      5250..5259
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             5280..5942
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     misc_feature    6021..6609
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6683..6916
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6988..7109
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7128..7263
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7301..7317)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7325..7341
                     /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(7349..7379)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7394..7415
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7703..8291)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8462..9322)
                     /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(9323..9427)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9901..9917
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagcgc
     2281 caccatggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaaaga
     2341 cgatgacgac aagcgcgggt ctagaatgct gctgcgcctg ctgctggcct gggtggccgc
     2401 ggtgcccgca ctgggccagg tcccctggac gccggagcct cgagccgcgt gcggccccag
     2461 cagctgctac gcgctctttc cccggcgccg cacattcctg gaagcttggc gggcgtgccg
     2521 cgaattgggg ggcaacctgg ccacaccgcg gaccccagag gaggcccagc gtgtggacag
     2581 cctggtgggg gtcgggccgg ccaacgggct gctatggatt gggttgcagc ggcaggctag
     2641 gcaatgccag ccgcagcgcc cactgcgggg cttcatatgg accacgggag accaggacac
     2701 cgccttcacc aactgggccc agccggctac ggaaggaccc tgcccagccc agcgctgtgc
     2761 agcccttgag gccagcggag agcatcgctg gctcgaaggc tcgtgcacac tggctgtcga
     2821 tggctacctc tgccagtttg gttttgaggg tgcctgccct gccttgccgc ttgaggtggg
     2881 tcaggccggt cccgctgtct acaccacacc cttcaacctg gtttccagcg agttcgaatg
     2941 gctgcccttt ggctccgtgg cagctgtgca gtgccaagct ggcaggggag cttctctgct
     3001 gtgcgtgaaa cagccttcag gtggcgtggg ctggtcccag actggcccgc tgtgcccagg
     3061 gactggctgt ggtcctgaca atgggggttg cgaacatgag tgtgtggaag aggtggacgg
     3121 tgctgtgtcc tgccgctgca gtgaaggctt ccgtctagca gcagatgggc acagttgtga
     3181 agacccctgt gcccaggccc cctgtgagca gcagtgtgaa cctggagggc cacaaggcta
     3241 tagctgccac tgtcgccttg gcttccggcc agctgaggat gatccacacc gctgcgtgga
     3301 cacggatgag tgccagattg ctggtgtgtg ccagcagatg tgtgtcaact atgttggtgg
     3361 ctttgagtgt tactgcagcg agggtcacga gcttgaggca gatggtatca gctgtagccc
     3421 tgcaggagcc atgggtgccc aggcttccca ggatctcaga gatgagttgc tggatgatgg
     3481 agaagaaggg gaggatgaag aggagccctg ggaggacttt gatggcacct ggacagagga
     3541 acaggggatc ctatggctgg cacctacaca tccacctgac tttggcctgc cctataggcc
     3601 caacttccca caggatggag agcctcagag attgcacctg gagcctacct ggccaccccc
     3661 acttagtgcc cccaggggcc cctaccactc ctcagtggtg tctgccacac ggcccatggt
     3721 gatctctgcc actcgaccca cactaccttc tgcccacaag acctctgtta tttcagctac
     3781 acgcccaccc ctgagccctg tccacccacc tgccatggcc cctgccacac ctccagctgt
     3841 gttctctgag caccagatcc ccaaaatcaa ggccaattat ccagacctgc cttttggcca
     3901 caagcctggg ataacctcgg ccactcaccc agcacggtct cctccgtacc agccccccat
     3961 tatctcaacc aactatcccc aagtcttccc tccccaccag gcccctatgt ctccagatac
     4021 ccacactatc acttatttgc ctccagtccc ccctcacctt gatcctgggg ataccacttc
     4081 taaagcccat caacaccctt tgctcccaga tgctccaggt atcagaaccc aggcccccca
     4141 gctttctgtc tcagctctcc agccccctct tcctaccaac tccaggtctt ctgtccatga
     4201 aactcctgtg cctgctgcca accagccccc agccttccct tcttctcccc tcccccctca
     4261 gaggcccact aaccagacct catctatcag ccctacacat tcctattcca gagcccctct
     4321 agtcccaagg gaaggagttc ccagtcccaa atcagtgcca cagctgccct cggtgccctc
     4381 cacagcagct ccaacagccc tggcagagtc aggtcttgca ggccaaagcc aaagggatga
     4441 ccgctggctg ctggtggcac tcctggtgcc aacatgtgtc ttcttggtgg tgctgcttgc
     4501 cctgggcatt gtgtactgca ctcgctgtgg ctcccacgca cccaacaagc ggatcacgga
     4561 ctgctatcgc tgggtcacac atgctgggaa caagagctca acagaaccca tgccccccag
     4621 aggcagcctt acaggggtac agacctgtag aaccagtgtg tgaggatccg cggccgccta
     4681 attaattaag gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca
     4741 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga
     4801 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     4861 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     4921 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc
     4981 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct
     5041 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg
     5101 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc
     5161 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat
     5221 ccaagctgtg accggcgcct acgctagacg ccaccatgga gagcgacgag agcggcctgc
     5281 ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg
     5341 tgggcggcgg agagggcacc cccaagcagg gccgcatgac caacaagatg aagagcacca
     5401 aaggcgccct gaccttcagc ccctacctgc tgagccacgt gatgggctac ggcttctacc
     5461 acttcggcac ctaccccagc ggctacgaga accccttcct gcacgccatc aacaacggcg
     5521 gctacaccaa cacccgcatc gagaagtacg aggacggcgg cgtgctgcac gtgagcttca
     5581 gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc
     5641 ccgaggacag cgtgatcttc accgacaaga tcatccgcag caacgccacc gtggagcacc
     5701 tgcaccccat gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg
     5761 acggcggcta ctacagcttc gtggtggaca gccacatgca cttcaagagc gccatccacc
     5821 ccagcatcct gcagaacggg ggccccatgt tcgccttccg ccgcgtggag gagctgcaca
     5881 gcaacaccga gctgggcatc gtggagtacc agcacgcctt caagaccccc atcgccttcg
     5941 ccagatcccg cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca
     6001 ccggatctcg ctaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     6061 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     6121 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     6181 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     6241 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     6301 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     6361 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     6421 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     6481 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6541 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     6601 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6661 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg
     6721 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6781 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6841 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6901 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     6961 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     7021 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7081 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     7141 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     7201 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     7261 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt
     7321 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7381 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7441 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7501 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7561 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7621 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7681 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7741 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7801 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7861 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7921 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7981 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8041 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8101 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     8161 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8221 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8281 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8341 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8401 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8461 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8521 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8581 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8641 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8701 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     8761 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8821 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8881 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8941 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9001 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9061 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9121 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9181 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9241 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9301 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9361 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9421 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     9481 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     9541 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     9601 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     9661 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     9721 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     9781 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     9841 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     9901 gtaaaacgac ggccagtgcc aagctg
//
