LOCUS       V042720                 9473 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042720
VERSION     V042720
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"
     regulatory      2284..2293
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2293..2358
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2359..3822
                     /label="Acod1(NM_008392)"
                     /note="Acod1(NM_008392)"
                     /gene="Acod1"
     CDS             3880..4590
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     promoter        4638..4849
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4862..5130
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5163..5561
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5568..6156
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6230..6463
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6535..6656
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6675..6810
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6848..6864)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6872..6888
                     /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(6896..6926)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6941..6962
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7250..7838)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8009..8869)
                     /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(8870..8974)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9448..9464
                     /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 agagccacca tggactacaa agaccatgac ggtgattata aagatcatga catcgactac
     2341 aaggatgacg atgacaagat gatgctcaag tctgtcacag agagctttgc tggtatgatt
     2401 cacggcttga aagtgaacca cctgacagat ggtatcattc ggaggagcaa gaggatgatc
     2461 ctggattctc tgggcgttgg cttcctgggg acaggcacag aagtgttcca taaagtcacc
     2521 caatatagta aaatctacag ttccaacacc tccagcactg tttggggtcg accagacttc
     2581 aggctcccac cgacatatgc tgcttttgtt aatggtgttg ctgttcactc catggatttt
     2641 gatgacacat ggcaccctgc cacccaccct tctggggctg tcctacctgt cctcacagct
     2701 ctatcggaag ccctgcctca gactcccaag ttttctggcc tcgacctgct gctggcgttc
     2761 aacgttggta ttgaagtaca gggacgatta atgcacttct ccaaggaagc caaagacata
     2821 ccaaagagat tccaccctcc ctctgtggtg gggactctgg gaagtgctgc tgctgcgtcc
     2881 aagtttctgg ggctcagctt gacaaagtgc cgcgaggcat tggctattgc tgtttcccac
     2941 gcaggggcac ccatagcgaa cgctgccact cagactaagc cccttcatat tggcaatgca
     3001 gccaagcatg ggatggaagc cacgtttctg gcaatgctgg gcctccaagg aaacaaacag
     3061 atcttggacc tggggtcagg gttcggtgcc ttctatgcca actactcccc cgaagacctt
     3121 ccaagcctgg attctcacat ctggctgttg gaccagcagg atgtggcctt taagagcttc
     3181 ccggcacatc tggctaccca ctgggtggca gatgcagctg cagccgtgag aaagcacctt
     3241 gtgacaccag aaagagccct gttccctgct gaccacatcg agagaatcgt gctcaggatc
     3301 cctgacgtcc agtacgtaaa caggcccttc ccggactcag agcatgaagc ccgtcattct
     3361 ttccagtatg tggcctgtgc ctcgctgctc gacggtagca tcactgtccc atccttccac
     3421 agccagcagg tcaataggcc tcaggtgaga gagttgctca agaaggtgaa gctggagcat
     3481 cctcctgaca acccgccaag cttcgacacg ctatactgtg aaataagcat cactctaaag
     3541 gacgggacca ctttcaccga gcgctctgac accttctatg gtcactggag gaaaccactg
     3601 agccaggaag atctgcgcaa caagttccga gccaatgcct caaagatgct atgcagggac
     3661 acggtggaaa gccttataac ggtagtagaa aagctagaag acctagaaga ctgctctgtg
     3721 ctaaccagac ttctgaaagg accctctgtc caagatgaag cttcaaaact atccagcatg
     3781 tcctcattcg atcacacaac gttgcccagg tttaccaata tcggtggagg cggatcagga
     3841 ggcggtggat ctggtggagg cggatctgaa ttcgccacca tggtgagcaa gggcgaggag
     3901 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     3961 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     4021 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     4081 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     4141 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     4201 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     4261 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     4321 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     4381 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     4441 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     4501 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     4561 accggcggca tggacgagct gtacaagtag ggatccgcgg ccgcgaagga tctgcgatcg
     4621 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     4681 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     4741 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     4801 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg
     4861 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg
     4921 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta
     4981 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc
     5041 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt
     5101 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctaggcca
     5161 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     5221 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     5281 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     5341 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     5401 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     5461 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     5521 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agtcgacaat caacctctgg
     5581 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5641 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5701 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5761 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5821 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5881 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5941 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     6001 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6061 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6121 agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact
     6181 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6241 attcactccc aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac
     6301 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6361 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6421 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     6481 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     6541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6661 gtctggctct agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6721 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     6781 agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt
     6841 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6901 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6961 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7021 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7081 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7141 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7201 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7261 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7321 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7681 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7861 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7921 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7981 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8041 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8101 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8161 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8221 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8281 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8341 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8401 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8461 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8521 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8581 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8641 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8701 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8761 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8821 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8881 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8941 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9001 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     9061 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     9121 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     9181 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     9241 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     9301 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     9361 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     9421 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
