LOCUS       V032320                10169 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032320
VERSION     V032320
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             2287..2352
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2377..5094
                     /label="CTNNA1(NM_001903)"
                     /note="CTNNA1(NM_001903)"
                     /gene="CTNNA1"
     promoter        5139..5350
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5363..5631
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5658..6257
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    6264..6852
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6926..7159
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7231..7352
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7371..7506
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7544..7560)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7568..7584
                     /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(7592..7622)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7637..7658
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7946..8534)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8705..9565)
                     /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(9566..9670)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10144..10160
                     /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 agaatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     2341 gacgatgaca agtctagaga attcggatcc gccaccatga ctgctgtcca tgcaggcaac
     2401 ataaacttca agtgggatcc taaaagtcta gagatcagga ctctggcagt tgagagactg
     2461 ttggagcctc ttgttacaca ggttacaacc cttgtaaaca ccaatagtaa agggccctct
     2521 aataagaaga gaggtcgttc taagaaggcc catgttttgg ctgcatctgt tgaacaagca
     2581 actgagaatt tcttggagaa gggggataaa attgcgaagg agagccagtt tctcaaggag
     2641 gagcttgtgg ctgctgtaga agatgttcga aaacaaggtg atttgatgaa ggctgctgca
     2701 ggagagttcg cagatgatcc ctgctcttct gtgaagcgag gcaacatggt tcgggcagct
     2761 cgagctttgc tctctgctgt tacccggttg ctgattttgg ctgacatggc agatgtctac
     2821 aaattacttg ttcagctgaa agttgtggaa gatggtatct tgaagttgag gaatgctggc
     2881 aatgaacaag acttaggaat ccagtataaa gccctaaaac ctgaagtgga taagctgaac
     2941 attatggcag ccaaaagaca acaggaattg aaagatgttg gccatcgtga tcagatggct
     3001 gcagctagag gaatcctgca gaagaacgtt ccgatcctct atactgcatc ccaggcatgc
     3061 ctacagcacc ctgatgtcgc agcctataag gccaacaggg acctgatata caagcagctg
     3121 cagcaggcgg tcacaggcat ttccaatgca gcccaggcca ctgcctcaga cgatgcctca
     3181 cagcaccagg gtggaggagg aggagaactg gcatatgcac tcaataactt tgacaaacaa
     3241 atcattgtgg accccttgag cttcagcgag gagcgcttta ggccttccct ggaggagcgt
     3301 ctggaaagca tcattagtgg ggctgccttg atggccgact cgtcctgcac gcgtgatgac
     3361 cgtcgtgagc gaattgtggc agagtgtaat gctgtccgcc aggccctgca ggacctgctt
     3421 tcggagtaca tgggcaatgc tggacgtaaa gaaagaagtg atgcactcaa ttctgcaata
     3481 gataaaatga ccaagaagac cagggacttg cgtagacagc tccgcaaagc tgtcatggac
     3541 cacgtttcag attctttcct ggaaaccaat gttccacttt tggtattgat tgaagctgca
     3601 aagaatggaa atgagaaaga agttaaggag tatgcccaag ttttccgtga acatgccaac
     3661 aaattgattg aggttgccaa cttggcctgt tccatctcaa ataatgaaga aggtgtaaag
     3721 cttgttcgaa tgtctgcaag ccagttagaa gccctctgtc ctcaggttat taatgctgca
     3781 ctggctttag cagcaaaacc acagagtaaa ctggcccaag agaacatgga tctttttaaa
     3841 gaacaatggg aaaaacaagt ccgtgttctc acagatgctg tcgatgacat tacttccatt
     3901 gatgacttct tggctgtctc agagaatcac attttggaag atgtgaacaa atgtgtcatt
     3961 gctctccaag agaaggatgt ggatggcctg gaccgcacag ctggtgcaat tcgaggccgg
     4021 gcagcccggg tcattcacgt agtcacctca gagatggaca actatgagcc aggagtctac
     4081 acagagaagg ttctggaagc cactaagctg ctctccaaca cagtcatgcc acgttttact
     4141 gagcaagtag aagcagccgt ggaagccctc agctcggacc ctgcccagcc catggatgag
     4201 aatgagttta tcgatgcttc ccgcctggta tatgatggca tccgggacat caggaaagca
     4261 gtgctgatga taaggacccc tgaggagttg gatgactctg actttgagac agaagatttt
     4321 gatgtcagaa gcaggacgag cgtccagaca gaagacgatc agctgatagc tggccagagt
     4381 gcccgggcga tcatggctca gcttccccag gagcaaaaag cgaagattgc ggaacaggtg
     4441 gccagcttcc aggaagaaaa gagcaagctg gatgctgaag tgtccaaatg ggacgacagt
     4501 ggcaatgaca tcattgtgct ggccaagcag atgtgcatga ttatgatgga gatgacagac
     4561 tttacccgag gtaaaggacc actcaaaaat acatcggatg tcatcagtgc tgccaagaaa
     4621 attgctgagg caggatccag gatggacaag cttggccgca ccattgcaga ccattgcccc
     4681 gactcggctt gcaagcagga cctgctggcc tacctgcaac gcatcgccct ctactgccac
     4741 cagctgaaca tctgcagcaa ggtcaaggcc gaggtgcaga atctcggcgg ggagcttgtt
     4801 gtctctgggg tggacagcgc catgtccctg atccaggcag ccaagaactt gatgaatgct
     4861 gtggtgcaga cagtgaaggc atcctacgtc gcctctacca aataccaaaa gtcacagggt
     4921 atggcttccc tcaaccttcc tgctgtgtca tggaagatga aggcaccaga gaaaaagcca
     4981 ttggtgaaga gagagaaaca ggatgagaca cagaccaaga ttaaacgggc atctcagaag
     5041 aagcacgtga acccggtgca ggccctcagc gagttcaaag ctatggacag catctaagcg
     5101 gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     5161 agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg
     5221 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     5281 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     5341 cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac
     5401 ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc
     5461 ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc
     5521 ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct
     5581 tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg
     5641 accggcgcct acgctagatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     5701 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     5761 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     5821 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     5881 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     5941 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     6001 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     6061 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     6121 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     6181 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     6241 gcaagcccgg tgcctgagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     6301 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     6361 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     6421 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     6481 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     6541 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6601 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     6661 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6721 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6781 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     6841 ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     6901 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat
     6961 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7021 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     7081 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     7141 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     7201 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     7261 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     7321 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     7381 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     7441 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7501 gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg
     7561 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     7621 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     7681 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     7741 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     7801 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     7861 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     7921 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     7981 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     8041 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     8101 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     8161 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     8221 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     8281 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     8341 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     8401 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     8461 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8521 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8581 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     8641 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8701 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8761 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8821 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8881 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8941 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     9001 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     9061 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     9121 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     9181 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     9241 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     9301 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     9361 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     9421 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9481 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9541 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9601 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9661 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     9721 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     9781 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     9841 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     9901 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     9961 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
    10021 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
    10081 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
    10141 gttgtaaaac gacggccagt gccaagctg
//
