LOCUS       V034361                 9156 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034361
VERSION     V034361
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3271
                     /label="NUDT18(NM_024815)"
                     /note="NUDT18(NM_024815)"
                     /gene="NUDT18"
     promoter        3315..3526
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3539..3807
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3836..3845
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3866..4528
                     /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             4598..4651
                     /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             4652..5251
                     /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    5252..5839
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5913..6146
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6218..6339
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6358..6493
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6531..6547)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6555..6571
                     /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(6579..6609)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6624..6645
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6933..7521)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7692..8552)
                     /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(8553..8657)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9131..9147
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatggcctc ggagggcctg gcgggggcgc tggcttccgt
     2341 gctggctggc caggggtcca gcgtgcacag ctgcgactcg gcgccggccg gggagccgcc
     2401 ggcgcccgtg cggctgcgga agaacgtgtg ctacgtggtg ctggccgtgt tcctcagcga
     2461 gcaggatgag gtgctactga tccaggaggc caagagggag tgccgggggt cgtggtacct
     2521 gcctgcgggg agaatggagc caggggagac catcgtggag gcgctgcagc gggaggtgaa
     2581 ggaggaggcg gggctgcact gtgagcccga gacactgctg tccgtggagg agcggggccc
     2641 ctcctgggtc cgcttcgtgt tcctcgctcg ccccacaggt ggaattctca agacttccaa
     2701 ggaggccgat gcggagtccc tgcaggctgc ctggtaccca cggacctccc tgcccactcc
     2761 gctgcgagcc catgacatcc tgcacctggt tgaactagcc gcccagtatc gccagcaagc
     2821 caggcaccct ctcattctgc cccaagagct accctgtgat ctggtctgcc agcggctcgt
     2881 ggctaccttt accagcgccc agacagtgtg ggtgttagtg ggcacagtgg ggatgcctca
     2941 cttgcctgtc actgcctgtg gcctcgaccc tatggagcag aggggtggca tgaagatggc
     3001 cgtcctgcgg ctgctgcagg agtgtctgac cctgcaccac ttggtggtgg agatcaaggg
     3061 gttgcttgga ctgcagcacc tgggccgaga tcacagtgat ggcatctgtt tgaatgtgct
     3121 ggtgaccgtg gcttttcgga gcccagggat ccaggatgaa cccccaaaag ttcggggtga
     3181 gaacttctct tggtggaagg tgatggagga agacctgcaa agccagctcc tccagcggct
     3241 tcagggatcc tctgttgtcc cagtgaacag atagcggccg cgaaggatct gcgatcgctc
     3301 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     3361 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     3421 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     3481 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg
     3541 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt
     3601 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt
     3661 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag
     3721 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc
     3781 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac
     3841 catggagagc gacgagagcg gcctgcccgc catggagatc gagtgccgca tcaccggcac
     3901 cctgaacggc gtggagttcg agctggtggg cggcggagag ggcaccccca agcagggccg
     3961 catgaccaac aagatgaaga gcaccaaagg cgccctgacc ttcagcccct acctgctgag
     4021 ccacgtgatg ggctacggct tctaccactt cggcacctac cccagcggct acgagaaccc
     4081 cttcctgcac gccatcaaca acggcggcta caccaacacc cgcatcgaga agtacgagga
     4141 cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag gccggccgcg tgatcggcga
     4201 cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg atcttcaccg acaagatcat
     4261 ccgcagcaac gccaccgtgg agcacctgca ccccatgggc gataacgtgc tggtgggcag
     4321 cttcgcccgc accttcagcc tgcgcgacgg cggctactac agcttcgtgg tggacagcca
     4381 catgcacttc aagagcgcca tccaccccag catcctgcag aacgggggcc ccatgttcgc
     4441 cttccgccgc gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg agtaccagca
     4501 cgccttcaag acccccatcg ccttcgccag atcccgcgct cagtcgtcca attctgccgt
     4561 ggacggcacc gccggacccg gctccaccgg atctcgcgag ggcagaggaa gtcttctaac
     4621 atgcggtgac gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct
     4681 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     4741 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     4801 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4861 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4921 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4981 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     5041 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     5101 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     5161 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     5221 ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga
     5281 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5341 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5401 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5461 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5521 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5581 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5641 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5701 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5761 gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga gtcggatctc
     5821 cctttggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     5881 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa
     5941 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     6001 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6061 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6121 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6181 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     6241 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6301 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     6361 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6421 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6481 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg
     6541 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6601 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6661 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6721 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6781 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6841 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6901 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6961 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7021 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7081 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7141 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7201 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7261 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7321 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7381 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7441 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7501 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7561 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7621 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7681 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7741 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7801 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7861 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7921 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7981 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8041 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8101 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8161 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8221 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8281 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8341 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8401 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8461 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8521 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8581 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8641 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     8701 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     8761 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     8821 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     8881 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     8941 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     9001 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     9061 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     9121 tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
