LOCUS       V024740                 9288 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024740
VERSION     V024740
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..2359
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     CDS             2366..3403
                     /label="CYSLTR2(NM_001308465)"
                     /note="CYSLTR2(NM_001308465)"
                     /gene="CYSLTR2"
     promoter        3447..3658
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3671..3939
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3968..3977
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3998..4660
                     /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             4730..4783
                     /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             4784..5383
                     /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    5384..5971
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6045..6278
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6350..6471
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6490..6625
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6663..6679)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6687..6703
                     /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(6711..6741)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6756..6777
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7065..7653)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7824..8684)
                     /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(8685..8789)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9263..9279
                     /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 caccatggac tacaaggatg acgatgacaa ggattacaaa gacgacgatg ataaggacta
     2341 taaggatgat gacgacaaag aattcatgga gagaaaattt atgtccttgc aaccatccat
     2401 ctccgtatca gaaatggaac caaatggcac cttcagcaat aacaacagca ggaactgcac
     2461 aattgaaaac ttcaagagag aatttttccc aattgtatat ctgataatat ttttctgggg
     2521 agtcttggga aatgggttgt ccatatatgt tttcctgcag ccttataaga agtccacatc
     2581 tgtgaacgtt ttcatgctaa atctggccat ttcagatctc ctgttcataa gcacgcttcc
     2641 cttcagggct gactattatc ttagaggctc caattggata tttggagacc tggcctgcag
     2701 gattatgtct tattccttgt atgtcaacat gtacagcagt atttatttcc tgaccgtgct
     2761 gagtgttgtg cgtttcctgg caatggttca cccctttcgg cttctgcatg tcaccagcat
     2821 caggagtgcc tggatcctct gtgggatcat atggatcctt atcatggctt cctcaataat
     2881 gctcctggac agtggctctg agcagaacgg cagtgtcaca tcatgcttag agctgaatct
     2941 ctataaaatt gctaagctgc agaccatgaa ctatattgcc ttggtggtgg gctgcctgct
     3001 gccatttttc acactcagca tctgttatct gctgatcatt cgggttctgt taaaagtgga
     3061 ggtcccagaa tcggggctgc gggtttctca caggaaggca ctgaccacca tcatcatcac
     3121 cttgatcatc ttcttcttgt gtttcctgcc ctatcacaca ctgaggaccg tccacttgac
     3181 gacatggaaa gtgggtttat gcaaagacag actgcataaa gctttggtta tcacactggc
     3241 cttggcagca gccaatgcct gcttcaatcc tctgctctat tactttgctg gggagaattt
     3301 taaggacaga ctaaagtctg cactcagaaa aggccatcca cagaaggcaa agacaaagtg
     3361 tgttttccct gttagtgtgt ggttgagaaa ggaaacaaga gtataagcgg ccgcaaggat
     3421 ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     3481 gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg
     3541 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     3601 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct
     3661 gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca
     3721 tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc
     3781 cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg
     3841 gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta
     3901 cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac
     3961 gctagacgcc accatggaga gcgacgagag cggcctgccc gccatggaga tcgagtgccg
     4021 catcaccggc accctgaacg gcgtggagtt cgagctggtg ggcggcggag agggcacccc
     4081 caagcagggc cgcatgacca acaagatgaa gagcaccaaa ggcgccctga ccttcagccc
     4141 ctacctgctg agccacgtga tgggctacgg cttctaccac ttcggcacct accccagcgg
     4201 ctacgagaac cccttcctgc acgccatcaa caacggcggc tacaccaaca cccgcatcga
     4261 gaagtacgag gacggcggcg tgctgcacgt gagcttcagc taccgctacg aggccggccg
     4321 cgtgatcggc gacttcaagg tggtgggcac cggcttcccc gaggacagcg tgatcttcac
     4381 cgacaagatc atccgcagca acgccaccgt ggagcacctg caccccatgg gcgataacgt
     4441 gctggtgggc agcttcgccc gcaccttcag cctgcgcgac ggcggctact acagcttcgt
     4501 ggtggacagc cacatgcact tcaagagcgc catccacccc agcatcctgc agaacggggg
     4561 ccccatgttc gccttccgcc gcgtggagga gctgcacagc aacaccgagc tgggcatcgt
     4621 ggagtaccag cacgccttca agacccccat cgccttcgcc agatcccgcg ctcagtcgtc
     4681 caattctgcc gtggacggca ccgccggacc cggctccacc ggatctcgcg agggcagagg
     4741 aagtcttcta acatgcggtg acgtggagga gaatcccggc cctatgaccg agtacaagcc
     4801 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     4861 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     4921 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4981 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     5041 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     5101 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     5161 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5221 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     5281 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5341 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac ctctggatta
     5401 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5461 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5521 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5581 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5641 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5701 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5761 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     5821 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5881 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt ctccgccttc gccctcagac
     5941 gagtcggatc tccctttggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa
     6001 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     6061 ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     6121 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6181 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6241 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     6301 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     6361 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6421 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6481 gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6541 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6601 tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa ttcgtaatca
     6661 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6721 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6781 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6841 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6901 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6961 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7021 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7081 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7141 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7201 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7261 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7321 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7381 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7441 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7501 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7561 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7621 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7681 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7741 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7801 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7861 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7921 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7981 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8041 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8101 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8161 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8221 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8281 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8341 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8401 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8461 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8521 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8581 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8641 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8701 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8761 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8821 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     8881 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     8941 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     9001 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     9061 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     9121 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     9181 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     9241 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
