LOCUS       V044939                 9586 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044939
VERSION     V044939
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..4805
                     /label="KLHDC7A(NM_152375)"
                     /note="KLHDC7A(NM_152375)"
                     /gene="KLHDC7A"
     CDS             4812..4877
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             4887..4943
                     /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             4956..5675
                     /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    5682..6269
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6343..6576
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6648..6769
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6788..6923
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6961..6977)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6985..7001
                     /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(7009..7039)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7054..7075
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7363..7951)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8122..8982)
                     /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(8983..9087)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9561..9577
                     /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 caccatgttc cccagaggag cagaggccca ggactggcat ttggatatgc
     2521 agctgaccgg caaggtggtg ctgtcagccg ctgccctgct cctggtgact gtggcctaca
     2581 ggctgtacaa gtcgaggcct gccccagccc agcggtgggg tgggaatggc caggcagaag
     2641 ccaaggagga agcagagggc tcagggcagc ctgctgtaca ggaggcttct cctggggtgc
     2701 tcctgagggg gccaagacgt cggaggagca gcaagcgggc tgaagcacca cagggctgca
     2761 gctgtgagaa tccaagaggc ccctatgtcc tggtcacggg ggccacttcc acagacagga
     2821 agccccagag aaaaggctca ggtgaggagc ggggcgggca gggctcggac tctgagcagg
     2881 tgcctccttg ctgccccagc caggaaacca gaacagctgt tggcagtaac cctgaccctc
     2941 cccatttccc ccgcttgggc agcgaaccga agagctcccc agctggactc attgcagcag
     3001 ccgacggcag ctgtgccggt ggtgagcctt ctccatggca ggacagtaaa ccccgtgagc
     3061 atccaggact ggggcaacta gaacctcccc actgtcacta cgtggctccc ttgcaaggca
     3121 gcagtgacat gaaccagagc tgggtcttca cccgtgtgat aggggtcagc agagaagagg
     3181 ctggggctct cgaggctgcc tccgatgttg acctgaccct gcatcagcag gagggcgccc
     3241 ccaactcctc ctataccttc tcatccatag cccgcgtccg aatggaggag catttcatac
     3301 agaaggcgga gggggttgag ccccggctca agggcaaggt gtacgactac tatgtggaat
     3361 ctacctctca ggccatcttc cagggcaggc tggctcccag gacagcagcc ctgactgagg
     3421 ttccatcccc taggccaccg ccagggtccc tgggaacagg ggctgcctcg ggaggccaag
     3481 ccggtgacac aaagggtgca gccgaaagag ccgcctcccc gcagacaggg ccgtggccct
     3541 ccacccgagg cttcagccgg aaggagagcc ttctgcagat agcggagaac ccagagctgc
     3601 agctgcagcc agatggcttc cggctccccg ctccaccctg cccagacccg ggcgccctgc
     3661 ctggcttagg cagaagcagc cgggagcccc atgtgcagcc ggtggccggg accaatttct
     3721 tccatatccc gctcacccct gcttcagccc cacaggtccg cctggatctg ggcaattgct
     3781 atgaggtgct gaccttggcc aagaggcaga acctggaggc cctgaaagag gcggcctaca
     3841 aggtgatgag cgaaaactac ctgcaggtgc tgcgcagccc ggacatctac gggtgcctga
     3901 gcggggcaga gcgcgagctg atcctgcagc gccggctccg gggccgccag tacctggtgg
     3961 tggctgacgt gtgccccaag gaagactccg gcggcctctg ttgctatgac gatgagcagg
     4021 atgtctggcg cccgctggct cgcatgcccc ccgaggccgt gtcccggggc tgtgccatct
     4081 gcagtctctt caattatctc ttcgtggtgt ccggctgcca ggggcccggg caccagccct
     4141 ccagccgcgt cttctgctac aacccgctca cggggatctg gagcgaggtg tgcccgctga
     4201 accaggcccg gccgcactgc cggctggtgg ccctggacgg gcacctgtat gccatcggcg
     4261 gagagtgtct gaactcggtg gagcgttacg acccccgcct ggaccgctgg gactttgccc
     4321 cgccgctccc cagtgacacg ttcgccctgg cgcacacggc cacggtgcgt gccaaggaaa
     4381 tcttcgtcac cggcggctcg ctgcgcttcc tgctgttccg cttctctgcg caggagcagc
     4441 gctggtgggc cggccccacc gggggcagca aggaccgcac ggccgagatg gtggcggtca
     4501 acggctttct ctaccgcttt gacctcaacc gcagcctggg catcgccgtg taccgctgca
     4561 gcgccagcac ccggctctgg tacgagtgcg ccacgtaccg gacgccttac ccggatgcct
     4621 tccagtgcgc cgtggtggac aacctcatct actgcgtggg acgccggagc accctctgct
     4681 tcctagcaga ctctgtctca cccaggtttg tgcccaagga gctgcggagt ttcccggccc
     4741 cgcagggcac cctcctgccc accgtcctga ccttgcccac ccccgatttg cctcagacca
     4801 gggtcgtcga cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     4861 aggatgacga tgacaaggga agcggagcca ctaacttctc cctgttgaaa caagcagggg
     4921 atgtcgaaga gaatcccggg ccagaattcg ccaccatggt gagcaagggc gaggagctgt
     4981 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     5041 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     5101 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     5161 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     5221 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     5281 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     5341 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     5401 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     5461 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     5521 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     5581 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     5641 ggatcactct cggcatggac gagctgtaca agtaaggatc caatcaacct ctggattaca
     5701 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5761 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5821 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5881 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5941 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     6001 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     6061 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     6121 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     6181 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga
     6241 gtcggatctc cctttggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     6301 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     6361 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     6421 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6481 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6541 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6601 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6781 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6841 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6901 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg
     6961 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7021 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7201 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7261 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7321 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7381 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7441 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7501 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7561 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7621 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7681 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7741 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7801 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7861 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7921 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7981 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8041 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8101 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8161 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8221 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8281 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8341 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8401 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8461 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8521 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8581 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8641 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8701 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8761 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8821 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8881 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8941 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9001 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9061 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     9121 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     9181 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     9241 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     9301 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     9361 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     9421 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     9481 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     9541 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
