LOCUS       V024575                 9455 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024575
VERSION     V024575
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             2317..3498
                     /label="Esam(NM_027102)"
                     /note="Esam(NM_027102)"
                     /gene="Esam"
     CDS             3505..3570
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3614..3825
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3838..4106
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4135..4144
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4165..4827
                     /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             4897..4950
                     /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             4951..5550
                     /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    5551..6138
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6212..6445
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6517..6638
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6657..6792
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6830..6846)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6854..6870
                     /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(6878..6908)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6923..6944
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7232..7820)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7991..8851)
                     /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(8852..8956)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9430..9446
                     /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 ctgatacgaa ctcggaattc gccaccatga ttcttcaggc tggaaccccc
     2341 gagaccagct tgctgcgggt tttgttcctg ggactgagta cccttgctgc cttctcccga
     2401 gctcagatgg agttgcacgt gcccccgggc ctcaacaaat tggaagcggt agagggagaa
     2461 gaagtggtgc tccccgcctg gtacacgatg gcacgggagg agtcgtggtc ccacccccgg
     2521 gaggtgccca tcctgatctg gttcttggaa caagaaggga aggaaccaaa ccaggtgttg
     2581 tcttacatta atggagtcat gacaaataaa cctggaacag ccctggtcca ctctatctct
     2641 tcacggaatg tgtccctgcg cctgggggca ctccaggagg gagactctgg gacttaccgc
     2701 tgttctgtca atgtgcagaa tgatgaaggc aaaagtatag gccacagcat caaaagcata
     2761 gagctcaaag tgctggttcc tccagctcct ccatcctgta gtttacaggg tgtaccctat
     2821 gtcgggacca atgtgaccct gaactgcaag tccccaagga gtaaacctac tgctcagtac
     2881 cagtgggaga ggctggcccc atcctcccag gtcttctttg gaccagcctt agatgctgtt
     2941 cgtggatctt taaagctcac taacctttcc attgccatgt ctggagtcta tgtctgcaag
     3001 gctcaaaaca gagtgggctt tgccaagtgc aacgtgacct tggacgtgat gacagggtcc
     3061 aaggctgcag tggtcgctgg agcagttgtg ggcacttttg ttgggttggt gctgatagct
     3121 gggctggtcc tgttgtacca gcgccggagc aagaccttgg aagagctggc caatgatatc
     3181 aaggaagatg ccattgctcc ccggaccttg ccttggacca aaggctcaga cacaatctcc
     3241 aagaatggga cactttcttc ggtcacctca gcacgagctc tgcggccacc caaggctgct
     3301 cctccaagac ctggcacatt tactcccaca cccagtgtct ctagccaggc cctgtcctca
     3361 ccaagactgc ccagggtaga tgaaccccca cctcaggcag tgtccctgac cccaggtggg
     3421 gtttcttctt ctgctctgag ccgcatgggt gctgtgcctg tgatggtgcc tgcacagagt
     3481 caggctgggt ctcttgtggg atccgactac aaagaccatg acggtgatta taaagatcat
     3541 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc
     3601 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     3661 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     3721 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     3781 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     3841 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     3901 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     3961 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     4021 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     4081 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agacgccacc
     4141 atggagagcg acgagagcgg cctgcccgcc atggagatcg agtgccgcat caccggcacc
     4201 ctgaacggcg tggagttcga gctggtgggc ggcggagagg gcacccccaa gcagggccgc
     4261 atgaccaaca agatgaagag caccaaaggc gccctgacct tcagccccta cctgctgagc
     4321 cacgtgatgg gctacggctt ctaccacttc ggcacctacc ccagcggcta cgagaacccc
     4381 ttcctgcacg ccatcaacaa cggcggctac accaacaccc gcatcgagaa gtacgaggac
     4441 ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg ccggccgcgt gatcggcgac
     4501 ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga tcttcaccga caagatcatc
     4561 cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg ataacgtgct ggtgggcagc
     4621 ttcgcccgca ccttcagcct gcgcgacggc ggctactaca gcttcgtggt ggacagccac
     4681 atgcacttca agagcgccat ccaccccagc atcctgcaga acgggggccc catgttcgcc
     4741 ttccgccgcg tggaggagct gcacagcaac accgagctgg gcatcgtgga gtaccagcac
     4801 gccttcaaga cccccatcgc cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg
     4861 gacggcaccg ccggacccgg ctccaccgga tctcgcgagg gcagaggaag tcttctaaca
     4921 tgcggtgacg tggaggagaa tcccggccct atgaccgagt acaagcccac ggtgcgcctc
     4981 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5041 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     5101 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5161 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5221 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5281 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     5341 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5401 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5461 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5521 tggtgcatga cccgcaagcc cggtgcctga aatcaacctc tggattacaa aatttgtgaa
     5581 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5641 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5701 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5761 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5821 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     5881 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     5941 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6001 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6061 ctgccggctc tgcggcctct tccgcgtctc cgccttcgcc ctcagacgag tcggatctcc
     6121 ctttggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat
     6181 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa
     6241 taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6301 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6361 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6421 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6481 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     6541 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6601 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     6661 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6721 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6781 tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt
     6841 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6961 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7021 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7801 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7861 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7921 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7981 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8041 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8101 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8161 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8221 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8281 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8341 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8401 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8461 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8521 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8581 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8641 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8701 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8761 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8821 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8881 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8941 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     9001 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     9061 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     9121 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     9181 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     9241 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     9301 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     9361 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     9421 cacgacgttg taaaacgacg gccagtgcca agctg
//
