LOCUS       V044861                 9379 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044861
VERSION     V044861
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..4598
                     /label="KLHL11(NM_018143)"
                     /note="KLHL11(NM_018143)"
                     /gene="KLHL11"
     CDS             4605..4670
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             4680..4736
                     /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             4749..5468
                     /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    5475..6062
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6136..6369
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6441..6562
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6581..6716
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6754..6770)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6778..6794
                     /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(6802..6832)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6847..6868
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7156..7744)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7915..8775)
                     /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(8776..8880)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9354..9370
                     /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 caccatggcg gctgcggcag tggcggcggc ggcggcggcg gccgcggctg
     2521 catctcttca ggtactggag atggagagca tggagacggc cgccgccggc tcggcaggac
     2581 tggccgccga ggtccgaggc agcggcacgg tggacttcgg gcctgggccg gggatctctg
     2641 caatggaggc gagcgggggc gatccgggcc cagaagccga ggatttcgag tgcagctctc
     2701 actgctcaga gctgtcctgg cggcagaacg agcagcggcg ccagggcctc ttctgcgaca
     2761 ttaccctgtg cttcggcggg gctggaggcc gcgagttccg ggcccaccgc tcggtactgg
     2821 ctgccgccac cgagtacttc acgcccctgc tctcgggcca gttttccgag tcccgctcgg
     2881 gacgggtgga gatgcgcaag tggagctccg agccggggcc cgaacccgac acagtggaag
     2941 ccgtaatcga gtacatgtac accgggcgca tccgcgtcag cacgggcagc gtgcacgagg
     3001 tgctggagtt ggccgacagg ttcctactca ttcgtttaaa agaattttgt ggagaatttc
     3061 tcaagaaaaa acttcatctc tcaaattgtg tggcaattca tagcttagca cacatgtaca
     3121 ccctgagcca acttgctctg aaggctgctg atatgatacg gagaaatttc cacaaagtga
     3181 ttcaggatga agaattttat acgttacctt tccatctcat tagagactgg ctttcagatt
     3241 tggaaattac agttgattct gaagaggttc tctttgaaac cgttttgaaa tgggttcaga
     3301 gaaatgctga agagagagag agatactttg aagaactttt taaattgctc aggttgtccc
     3361 agatgaaacc tacctacctt actcgacatg tcaaaccaga gaggctggta gccaataatg
     3421 aagtttgtgt caagttggtc gctgacgcag tggagagaca tgctctgaga gctgagaata
     3481 tacaatctgg cacatgccag caccccactt ctcatgtgtc actattgcct cgttatgggc
     3541 aaaacatgga tgtgatcatg gttattggag gtgtgtcaga aggaggggac tatttaagtg
     3601 aatgtgtggg atactttgtt gatgaggaca gatgggtaaa tctgccacat attcataatc
     3661 acctcgatgg acatgctgtt gcagtaacag aatcctacgt gtatgttgct ggatcaatgg
     3721 agccagggtt tgctaaaact gtagaaaggt ataacccaaa tttgaataca tgggaacatg
     3781 tttgtagtct gatgacaaga aagcattctt ttggactaac agaagtcaaa gggaagctct
     3841 atagcattgg aggacatggc aactttagtc ctggttttaa agatgtgact gtttataatc
     3901 ctgagcttga taaatggcac aacttggaat cggcaccaaa gattcttcga gatgtcaaag
     3961 cactagccat tgaagaccgg tttgtataca ttgccgcccg cactcctgta gaccgggaca
     4021 ctgaagatgg attaaaggct gtaattactt gctatgatac agagactcga cagtggcaag
     4081 atgtggaatc tttgccgctt attgacaatt actgcttttt ccaaatgtct gtggtcaatt
     4141 caaactttta tcagacagca tcatgttgtc ccaagagtta ttgtttagaa aacgaagagg
     4201 cagtaagaaa aattgccagc caagtgtctg atgagatcct tgaaagcttg cctccagaag
     4261 tcctaagcat cgaaggagca gccatttgct attacaaaga tgatgtcttc attataggag
     4321 gctggaaaaa cagtgatgat attgataaac agtatcggaa agaagcctac cgatattgtg
     4381 cggagaggaa gaggtggatg cttcttcctc ctatgccaca acctcgttgt agagccactg
     4441 cttgtcacgt gaggatccca taccggtact tgcatggcac acagagatac cctatgcctc
     4501 aaaacctgat gtggcagaag gaccgcatca gacagatgca agagatacat cgtcacgccc
     4561 tgaacatgag gcgagtgcca agctctcaga ttgaatgcgt cgacgactac aaagaccatg
     4621 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag ggaagcggag
     4681 ccactaactt ctccctgttg aaacaagcag gggatgtcga agagaatccc gggccagaat
     4741 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     4801 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     4861 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     4921 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     4981 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     5041 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     5101 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     5161 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     5221 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     5281 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     5341 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     5401 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     5461 acaagtaagg atccaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     5521 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     5581 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     5641 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     5701 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     5761 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     5821 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     5881 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     5941 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     6001 ctcttccgcg tctccgcctt cgccctcaga cgagtcggat ctccctttgg ccgcctcccc
     6061 gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     6121 aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat ctgctttttg
     6181 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     6241 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     6301 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     6361 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     6421 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     6481 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6541 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     6601 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6661 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga
     6721 cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6781 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6841 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6901 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6961 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     7021 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     7081 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7561 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7801 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7861 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7921 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7981 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     8041 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8101 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8161 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8221 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8281 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8341 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8401 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8461 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8521 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8581 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8641 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8701 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8761 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8821 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8881 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     8941 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     9001 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     9061 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     9121 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     9181 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     9241 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     9301 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     9361 gacggccagt gccaagctg
//
