LOCUS       V030268                 8596 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030268
VERSION     V030268
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..3249
                     /label="BRCC3(NM_024332)"
                     /note="BRCC3(NM_024332)"
                     /gene="BRCC3"
     CDS             3256..3321
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3365..3576
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3589..3857
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3890..4684
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    4691..5279
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5353..5586
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5658..5779
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5798..5933
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5971..5987)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5995..6011
                     /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(6019..6049)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6064..6085
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6373..6961)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7132..7992)
                     /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(7993..8097)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8571..8587
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggcggtg caggtggtgc aggcggtgca ggcggttcat
     2341 ctcgagtctg acgctttcct cgtttgtctc aaccacgctc tgagcacaga gaaggaggaa
     2401 gtaatggggc tgtgcatagg ggagttgaac gatgatacaa ggagtgactc caaatttgca
     2461 tatactggaa ctgaaatgcg cacagttgct gaaaaggttg atgccgtcag aattgttcac
     2521 attcattctg tcatcatctt acgacgttct gataagagga aggaccgagt agaaatttct
     2581 ccagagcagc tgtctgcagc ttcaacagag gcagagaggt tggctgaact gacaggccgc
     2641 cccatgagag ttgtgggctg gtatcattcc catcctcata taactgtttg gccttcacat
     2701 gttgatgttc gcacacaagc catgtaccag atgatggatc aaggctttgt aggacttatt
     2761 ttttcctgtt tcatagaaga taagaacaca aagactggcc gggtactcta cacttgcttc
     2821 caatccatac aggcccaaaa gagttcagag tcccttcatg gtccacgaga cttctggagc
     2881 tccagccagc acatctccat tgagggccag aaggaagagg aaaggtatga gagaatcgaa
     2941 atcccaatcc atattgtacc tcatgtcact atcgggaaag tgtgccttga atcagcagta
     3001 gagctgccca agatcctgtg ccaggaggag caggatgcgt ataggaggat ccacagcctt
     3061 acacatctgg actcagtaac caagatccat aatggctcag tgtttaccaa gaatctgtgc
     3121 agtcagatgt cggcagtcag cgggcctctc ctacagtggt tggaggacag actggagcaa
     3181 aaccaacagc atttgcagga attacaacaa gaaaaggaag agcttatgca agaactttct
     3241 tctctagaac tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3301 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     3361 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     3421 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     3481 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     3541 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     3601 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     3661 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     3721 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     3781 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     3841 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc taggccacca tggctgaaca
     3901 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3961 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4021 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4081 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4141 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4201 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4261 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4321 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4381 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4441 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4501 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4561 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4621 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4681 ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     4741 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     4801 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     4861 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     4921 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     4981 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5041 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     5101 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5161 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5221 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5281 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     5341 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt
     5401 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5461 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5521 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     5581 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     5641 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5701 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5761 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag
     5821 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5881 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     5941 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6001 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6061 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6121 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6181 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6781 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7021 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7081 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7141 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7201 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7261 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7321 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7381 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7441 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7501 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7561 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7621 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7681 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7741 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7801 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7861 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7921 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7981 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8041 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8101 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     8161 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     8221 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     8281 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     8341 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     8401 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     8461 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     8521 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     8581 ggccagtgcc aagctg
//
