LOCUS       V033052                 8071 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033052
VERSION     V033052
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..2758
                     /label="Id1(NM_001355113)"
                     /note="Id1(NM_001355113)"
                     /gene="Id1"
     CDS             2765..2830
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        2874..3085
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3098..3366
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3395..3404
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3425..4087
                     /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    4166..4754
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             4828..5061
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5133..5254
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5273..5408
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5446..5462)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5470..5486
                     /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(5494..5524)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5539..5560
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5848..6436)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6607..7467)
                     /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(7468..7572)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8046..8062
                     /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 gccaccatga aggtcgccag tggcagtgcc gcagccgctg caggccctag
     2341 ctgttcgctg aaggcgggca ggacagcggg cgaggtggta cttggtctgt cggagcaaag
     2401 cgtggccatc tcgcgctgcg ctgggacgcg cctgcccgcc ttgctggacg agcagcaggt
     2461 gaacgtcctg ctctacgaca tgaacggctg ctactcacgc ctcaaggagc tggtgcccac
     2521 cctgccccag aaccgcaaag tgagcaaggt ggagatcctg cagcatgtaa tcgactacat
     2581 cagggacctg cagctggagc tgaactcgga gtctgaagtc gggaccaccg gaggccgggg
     2641 actgcctgtc cgcgccccgc tcagcaccct gaacggcgag atcagtgcct tggcggccga
     2701 ggtgaggcgg catgtgttcc agccgacgat cgcatcttgt gtcgctgagg cggcgcacgg
     2761 atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2821 cgatgacaag tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga
     2881 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
     2941 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     3001 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     3061 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     3121 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     3181 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     3241 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     3301 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     3361 cagatccaag ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg
     3421 cctgcccgcc atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga
     3481 gctggtgggc ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag
     3541 caccaaaggc gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt
     3601 ctaccacttc ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa
     3661 cggcggctac accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag
     3721 cttcagctac cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg
     3781 cttccccgag gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga
     3841 gcacctgcac cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct
     3901 gcgcgacggc ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat
     3961 ccaccccagc atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct
     4021 gcacagcaac accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc
     4081 cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg
     4141 ctccaccgga tctcgctaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     4201 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4261 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     4321 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     4381 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     4441 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     4501 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     4561 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     4621 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4681 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4741 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     4801 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     4861 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     4921 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     4981 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5041 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     5101 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     5161 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5221 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     5281 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5341 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5401 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     5461 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5521 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5581 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5641 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5701 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5761 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5821 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5881 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5941 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6001 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6061 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6121 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6181 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6241 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6301 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6361 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6421 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6481 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6541 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6601 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6661 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6721 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6781 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6841 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6901 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6961 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7021 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7081 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7141 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7201 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7261 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7321 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7381 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7441 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7501 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7561 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7621 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     7681 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     7741 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     7801 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     7861 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     7921 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     7981 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     8041 acgttgtaaa acgacggcca gtgccaagct g
//
