LOCUS       V044884                 9130 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044884
VERSION     V044884
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"
     enhancer        1933..2236
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2237..2440
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2475..4349
                     /label="KLHL24(NM_001349413)"
                     /note="KLHL24(NM_001349413)"
                     /gene="KLHL24"
     CDS             4356..4421
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             4431..4487
                     /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 porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             4500..5219
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     misc_feature    5226..5813
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5887..6120
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6192..6313
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6332..6467
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6505..6521)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6529..6545
                     /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(6553..6583)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6598..6619
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6907..7495)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7666..8526)
                     /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(8527..8631)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9105..9121
                     /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 gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1981 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     2041 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2101 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2161 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2221 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2281 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2341 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     2401 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     2461 ccgctagcgc caccatggta ctaatattgg gacgcagact aaacagagag gatcttgggg
     2521 tgcgtgattc cccagcaact aagcgaaaag tttttgaaat ggaccccaaa tctctgacag
     2581 gtcatgagtt ttttgacttc tcttcaggat catcccatgc cgaaaacata ctccagatat
     2641 ttaatgaatt tcgtgatagc cgcttattca cagatgttat catttgtgtg gaaggaaaag
     2701 aatttccttg ccatagagct gtgctctcag cctgtagcag ctacttcaga gctatgtttt
     2761 gtaatgacca cagggaaagc cgagaaatgt tggttgagat caatggtatt ttagctgaag
     2821 ctatggaatg ttttttgcag tatgtttata ctggaaaggt gaagatcact acagagaatg
     2881 tacagtatct ctttgagaca tcaagcctct ttcagattag tgttctccgt gatgcatgtg
     2941 ccaagttctt ggaggagcaa cttgatcctt gtaattgctt aggaatccag cgctttgctg
     3001 atacccattc actcaaaaca ctcttcacaa aatgcaaaaa ttttgcgtta cagacttttg
     3061 aggatgtatc ccagcacgaa gaatttcttg agcttgacaa agatgaactt attgattata
     3121 tttgtagtga tgaacttgtt attggtaaag aggagatggt ttttgaagcc gtcatgcgtt
     3181 gggtctatcg tgccgttgat ctgagaagac cactgttaca cgagctcctg acacatgtga
     3241 gactccctct gttgcatccc aactactttg ttcaaacagt tgaagtggac caattgatcc
     3301 agaattctcc tgagtgttat cagttgttgc atgaagcaag acggtaccac atacttggga
     3361 atgaaatgat gtccccaagg actaggccac gcaggtccac tggctattct gaggtgatag
     3421 ttgtcgttgg aggatgtgag cgagttggag gatttaatct tccatacact gagtgctacg
     3481 atcctgtaac aggagaatgg aagtctttgg ctaagcttcc agaatttacc aaatcagagt
     3541 atgcagtctg tgctctaagg aatgacattc ttgtttcagg tggaagaatc aacagccgtg
     3601 atgtctggat ttataactca cagttaaata tttggatcag agttgcctct ctcaataaag
     3661 gcagatggcg tcacaaaatg gctgtcctcc ttggtaaagg gtttcttaaa acttctatac
     3721 taggccagtc gtggtggctc acacctgtaa tcccagcact ttgggaggcc aaggtatatg
     3781 ttgtcggtgg ctatgatggg caaaacagac ttagcagcgt agaatgttat gattcctttt
     3841 caaatcgatg gactgaagtt gctcccctta aggaagccgt gagttctcct gcagtgacta
     3901 gctgtgtagg caaactgttt gtgattggtg gaggacctga tgataatact tgttctgata
     3961 aggttcaatc ttatgatcca gaaaccaatt cttggctact tcgtgcagct atcccaattg
     4021 ccaaaaggtg tataacagct gtatccctaa acaacctgat ctatgttgcc ggtggactga
     4081 ccaaggcaat atactgttac gatccagttg aagattactg gatgcacgta cagaatacat
     4141 tcagccgtca ggaaaactgt ggtatgtctg tgtgtaatgg taaaatatat atcctgggcg
     4201 gaagacggga aaatggagaa gccacagaca ctattctctg ttatgatcct gcaacaagta
     4261 tcatcacagg ggtagctgca atgcccaggc cagtgtccta tcatggctgt gtgactattc
     4321 atagatacaa tgagaaatgc tttaaactcg tcgacgacta caaagaccat gacggtgatt
     4381 ataaagatca tgacatcgac tacaaggatg acgatgacaa gggaagcgga gccactaact
     4441 tctccctgtt gaaacaagca ggggatgtcg aagagaatcc cgggccagaa ttcgccacca
     4501 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     4561 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     4621 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     4681 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     4741 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     4801 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     4861 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     4921 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     4981 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     5041 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     5101 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     5161 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
     5221 gatccaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5281 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5341 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5401 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5461 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5521 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5581 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     5641 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5701 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5761 gtctccgcct tcgccctcag acgagtcgga tctccctttg gccgcctccc cgcctggtac
     5821 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     5881 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg
     5941 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6001 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6061 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6121 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     6181 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6241 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6301 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc
     6361 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6421 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag
     6481 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6541 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6601 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6661 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6721 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6781 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6841 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6901 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6961 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7021 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7081 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7141 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7201 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7261 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7321 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7381 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7441 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7501 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7561 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7621 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7681 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7741 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7801 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7861 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7921 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7981 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8041 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8101 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8161 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8221 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8281 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8341 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8401 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8461 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8521 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8581 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8641 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8701 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     8761 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     8821 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     8881 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     8941 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     9001 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     9061 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     9121 tgccaagctg
//
