LOCUS       V019296                 9447 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019296
VERSION     V019296
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"
     CDS             2297..3532
                     /label="ASS1(NM_000050)"
                     /note="ASS1(NM_000050)"
                     /gene="ASS1"
     CDS             3533..3556
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        3606..3817
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3830..4098
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4127..4136
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4157..4819
                     /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"
     CDS             4889..4942
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4943..5542
                     /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    5543..6130
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6204..6437
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6509..6630
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6649..6784
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6822..6838)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6846..6862
                     /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(6870..6900)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6915..6936
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7224..7812)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7983..8843)
                     /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(8844..8948)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9422..9438
                     /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 attctagagc
     2281 tagcgaattc gccaccatgt ccagcaaagg ctccgtggtt ctggcctaca gtggcggcct
     2341 ggacacctcg tgcatcctcg tgtggctgaa ggaacaaggc tatgacgtca ttgcctatct
     2401 ggccaacatt ggccagaagg aagacttcga ggaagccagg aagaaggcac tgaagcttgg
     2461 ggccaaaaag gtgttcattg aggatgtcag cagggagttt gtggaggagt tcatctggcc
     2521 ggccatccag tccagcgcac tgtatgagga ccgctacctc ctgggcacct ctcttgccag
     2581 gccctgcatc gcccgcaaac aagtggaaat cgcccagcgg gagggggcca agtatgtgtc
     2641 ccacggcgcc acaggaaagg ggaacgatca ggtccggttt gagctcagct gctactcact
     2701 ggccccccag ataaaggtca ttgctccctg gaggatgcct gaattctaca accggttcaa
     2761 gggccgcaat gacctgatgg agtacgcaaa gcaacacggg attcccatcc cggtcactcc
     2821 caagaacccg tggagcatgg atgagaacct catgcacatc agctacgagg ctggaatcct
     2881 ggagaacccc aagaaccaag cgcctccagg tctctacacg aagacccagg acccagccaa
     2941 agcccccaac acccctgaca ttctcgagat cgagttcaaa aaaggggtcc ctgtgaaggt
     3001 gaccaacgtc aaggatggca ccacccacca gacctccttg gagctcttca tgtacctgaa
     3061 cgaagtcgcg ggcaagcatg gcgtgggccg tattgacatc gtggagaacc gcttcattgg
     3121 aatgaagtcc cgaggtatct acgagacccc agcaggcacc atcctttacc atgctcattt
     3181 agacatcgag gccttcacca tggaccggga agtgcgcaaa atcaaacaag gcctgggctt
     3241 gaaatttgct gagctggtgt ataccggttt ctggcacagc cctgagtgtg aatttgtccg
     3301 ccactgcatc gccaagtccc aggagcgagt ggaagggaaa gtgcaggtgt ccgtcctcaa
     3361 gggccaggtg tacatcctcg gccgggagtc cccactgtct ctctacaatg aggagctggt
     3421 gagcatgaac gtgcagggtg attatgagcc aactgatgcc accgggttca tcaacatcaa
     3481 ttccctcagg ctgaaggaat atcatcgtct ccagagcaag gtcactgcca aagattacaa
     3541 ggatgacgac gataagtgag gatccgcggc cgcaaggatc tgcgatcgct ccggtgcccg
     3601 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     3661 ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     3721 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     3781 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat
     3841 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt
     3901 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa
     3961 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg
     4021 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt
     4081 tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagacgcca ccatggagag
     4141 cgacgagagc ggcctgcccg ccatggagat cgagtgccgc atcaccggca ccctgaacgg
     4201 cgtggagttc gagctggtgg gcggcggaga gggcaccccc aagcagggcc gcatgaccaa
     4261 caagatgaag agcaccaaag gcgccctgac cttcagcccc tacctgctga gccacgtgat
     4321 gggctacggc ttctaccact tcggcaccta ccccagcggc tacgagaacc ccttcctgca
     4381 cgccatcaac aacggcggct acaccaacac ccgcatcgag aagtacgagg acggcggcgt
     4441 gctgcacgtg agcttcagct accgctacga ggccggccgc gtgatcggcg acttcaaggt
     4501 ggtgggcacc ggcttccccg aggacagcgt gatcttcacc gacaagatca tccgcagcaa
     4561 cgccaccgtg gagcacctgc accccatggg cgataacgtg ctggtgggca gcttcgcccg
     4621 caccttcagc ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc acatgcactt
     4681 caagagcgcc atccacccca gcatcctgca gaacgggggc cccatgttcg ccttccgccg
     4741 cgtggaggag ctgcacagca acaccgagct gggcatcgtg gagtaccagc acgccttcaa
     4801 gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc aattctgccg tggacggcac
     4861 cgccggaccc ggctccaccg gatctcgcga gggcagagga agtcttctaa catgcggtga
     4921 cgtggaggag aatcccggcc ctatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4981 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     5041 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     5101 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     5161 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     5221 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5281 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5341 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5401 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5461 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5521 gacccgcaag cccggtgcct gaaatcaacc tctggattac aaaatttgtg aaagattgac
     5581 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5641 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5701 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5761 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5821 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5881 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     5941 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     6001 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     6061 tctgcggcct cttccgcgtc tccgccttcg ccctcagacg agtcggatct ccctttggcc
     6121 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     6181 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct
     6241 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6301 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6361 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6421 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6481 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     6541 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6601 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     6661 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6721 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6781 ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg
     6841 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6901 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6961 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7021 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7621 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7861 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7921 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7981 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8041 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8101 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8161 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8221 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8281 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8341 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8401 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8461 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8521 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8581 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8641 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8701 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8761 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8821 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8881 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8941 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     9001 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     9061 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     9121 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     9181 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     9241 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     9301 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     9361 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     9421 tgtaaaacga cggccagtgc caagctg
//
