LOCUS       V046687                 8629 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046687
VERSION     V046687
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..3848
                     /label="KBTBD13(NM_001101362)"
                     /note="KBTBD13(NM_001101362)"
                     /gene="KBTBD13"
     CDS             3855..3920
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             3930..3986
                     /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             3999..4718
                     /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    4725..5312
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5386..5619
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5691..5812
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5831..5966
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6004..6020)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6028..6044
                     /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(6052..6082)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6097..6118
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6406..6994)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7165..8025)
                     /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(8026..8130)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8604..8620
                     /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 caccatggca cggggtccac agaccctggt gcaggtgtgg gtgggcggcc
     2521 agctcttcca agccgaccgc gccctgctgg tggagcactg tggcttcttc cgaggcctct
     2581 tccgctccgg catgcgggag acccgcgcag cagaggtgcg cctgggcgtt ctgagcgcgg
     2641 gaggtttccg cgccacgctg caggtgctgc gcggcgaccg gccggcgctg gcggcggagg
     2701 acgagctgct gcaggccgtg gagtgcgccg ccttcctcca ggcgccggcg ctggctcgct
     2761 ttctggagca caacctcacg tcggacaact gcgcattgct gtgcgacgcg gccgccgcct
     2821 tcggcctgcg cgacgtgttc cacagtgccg cgctcttcat ctgcgacggc gagcgcgagc
     2881 tggcggccga actggcgctg cctgaggccc gcgcctacgt ggcggccctg cggcccagca
     2941 gctacgcggc cgtgagcacg cacacgcccg cgcccggctt cctggaggac gcctcgcgca
     3001 cgctgtgtta cctggacgag gaagaggacg cgtggcgcac gctggctgcg ctgcccctgg
     3061 aggccagcac gttgctggcc ggggtggcca cgctgggcaa caagctttac atcgtggggg
     3121 gcgtgcgcgg cgccagcaag gaggtggtag agctgggctt ctgctacgac cccgacggcg
     3181 gcacgtggca cgagttcccc agcccgcacc agccgcgcta tgacacagcg ctggccggct
     3241 tcgacggccg cctctacgcc atcggcggcg aattccagag gacgcccatc agctccgtgg
     3301 agcgctacga cccagccgcg ggctgctgga gtttcgtggc cgacctgccg cagccggccg
     3361 ccggcgtgcc ctgcgcccag gcttgtggcc gtctcttcgt gtgcctgtgg cggccggccg
     3421 acaccaccgc cgtggtggag tacgcagtgc ggaccgacgc gtggctgcca gtggccgagc
     3481 tgcggcgtcc gcagagctat ggccactgca tggtggccca ccgcgacagc ctctatgtgg
     3541 tgcgcaacgg accttccgac gacttcctgc actgcgccat cgactgtctc aacctggcca
     3601 cgggccagtg gacggcgctg cccggccagt tcgtcaacag caagggagcg ctcttcacgg
     3661 ccgtggtgcg cggtgacacc gtctatacgg tcaaccgcat gttcacgctg ctctacgcca
     3721 tcgagggcgg cacctggcgg ctgctcaggg agaaagccgg cttcccgcgg cccggctcct
     3781 tgcagacctt tctcctaagg ctgcctcctg gcgctcctgg gcctgtgact tcgacaacgg
     3841 cagaactggt cgacgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     3901 acaaggatga cgatgacaag ggaagcggag ccactaactt ctccctgttg aaacaagcag
     3961 gggatgtcga agagaatccc gggccagaat tcgccaccat ggtgagcaag ggcgaggagc
     4021 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     4081 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     4141 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     4201 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     4261 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     4321 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     4381 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     4441 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     4501 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     4561 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     4621 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     4681 ccgggatcac tctcggcatg gacgagctgt acaagtaagg atccaatcaa cctctggatt
     4741 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     4801 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     4861 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     4921 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     4981 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     5041 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     5101 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     5161 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     5221 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tctccgcctt cgccctcaga
     5281 cgagtcggat ctccctttgg ccgcctcccc gcctggtacc tttaagacca atgacttaca
     5341 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     5401 actcccaacg aaaataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     5461 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     5521 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     5581 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     5641 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     5701 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5761 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5821 ggctctagct atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5881 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     5941 gtagtgagga ggcttttttg gaggcctaga cttttgcaga gacggcccaa attcgtaatc
     6001 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6061 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6121 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6181 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6241 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6301 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6361 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6421 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6481 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6541 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6601 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6661 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6721 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6781 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6841 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6901 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6961 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7021 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7081 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7141 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7201 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7261 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7321 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7381 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7441 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7501 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7561 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7621 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7681 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7741 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7801 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7861 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7921 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7981 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8041 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8101 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8161 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     8221 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     8281 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     8341 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     8401 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     8461 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     8521 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     8581 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagctg
//
