LOCUS       V044597                 9310 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044597
VERSION     V044597
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             2302..3009
                     /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"
     CDS             3040..4359
                     /label="Pla2g7(NM_013737)"
                     /note="Pla2g7(NM_013737)"
                     /gene="Pla2g7"
     CDS             4366..4431
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4475..4686
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4699..4967
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5000..5398
                     /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    5405..5993
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6067..6300
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6372..6493
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6512..6647
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6685..6701)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6709..6725
                     /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(6733..6763)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6778..6799
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7087..7675)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7846..8706)
                     /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(8707..8811)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9285..9301
                     /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 agagctagcg aattcgccac catggtgagc aagggcgagg aggataacat ggccatcatc
     2341 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     2401 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     2461 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     2521 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     2581 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     2641 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     2701 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     2761 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     2821 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     2881 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     2941 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     3001 ctgtacaagg gaggtggagg tagtggagga ggcggatcaa tggtaccact caaactgcag
     3061 gcgcttttct gcctcctctg ctgcctccca tgggtccatc cttttcactg gcaagacaca
     3121 tcttcttttg acttcaggcc gtcagtaatg tttcacaagc tccaatcggt gatgtctgct
     3181 gccggctctg gccatagtaa aatccccaaa ggaaatggat cgtaccccgt cggttgtaca
     3241 gatctgatgt tcggttatgg gaatgagagc gtcttcgtgc gtttgtacta cccagctcaa
     3301 gatcaaggtc gcctcgacac tgtttggatc ccaaacaaag aatatttttt gggtcttagt
     3361 atatttcttg gaacacccag tattgtaggc aatattttac acctcttata tggttctctg
     3421 acaactcctg caagctggaa ttctcctttg aggactggag aaaaataccc actcattgtc
     3481 ttttctcatg gtctcggagc cttcaggacg atttattctg ctattggcat tggcttggca
     3541 tctaatgggt ttatagtggc cactgtcgaa cacagagaca gatctgcatc ggcaacttac
     3601 ttttttgaag accaggtggc tgcaaaagtg gaaaacaggt cttggcttta cctgagaaaa
     3661 gtaaaacaag aggagtcgga aagtgtccgg aaagaacagg ttcagcaaag agcaatagaa
     3721 tgttcccggg ctctcagtgc gattcttgac attgaacatg gagacccaaa agagaatgta
     3781 ctaggttcag cttttgacat gaaacagctg aaggatgcta ttgatgagac taaaatagct
     3841 ttgatgggac attcttttgg aggagcaaca gttcttcaag cccttagtga ggaccagaga
     3901 ttcagatgtg gagttgctct tgatccatgg atgtatccgg tgaacgaaga gctgtactcc
     3961 agaaccctcc agcctctcct ctttatcaac tctgccaaat tccagactcc aaaggacatc
     4021 gcaaaaatga aaaagttcta ccagcctgac aaggaaagga aaatgattac aatcaagggc
     4081 tcagtgcacc agaactttga cgactttact tttgtaactg gcaaaataat tggaaacaag
     4141 ctgacactga aaggagaaat cgattccaga gtagccatcg acctcaccaa caaagcttcg
     4201 atggctttct tacaaaagca tttagggctt cagaaagact ttgatcagtg ggaccctctg
     4261 gtggaaggag atgatgagaa cctgattcct gggtcaccct ttgacgcagt cacccaggtc
     4321 ccggctcagc aacactctcc aggatcacag acccagaatg gatccgacta caaagaccat
     4381 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg
     4441 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     4501 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     4561 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     4621 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     4681 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     4741 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     4801 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     4861 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     4921 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     4981 gcgcctacgc taggccacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     5041 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     5101 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     5161 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     5221 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     5281 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     5341 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt
     5401 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5461 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5521 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5581 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5641 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5701 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5761 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     5821 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5881 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5941 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac
     6001 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     6061 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg
     6121 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6181 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6241 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6301 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     6361 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6421 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6481 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc
     6541 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6601 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag
     6661 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6721 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6781 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6841 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6901 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6961 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7021 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7081 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7141 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7201 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7261 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7321 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7381 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7441 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7501 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7561 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7621 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7681 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7741 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7801 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7861 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7921 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7981 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8041 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8101 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8161 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8221 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8281 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8341 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8401 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8461 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8521 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8581 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8641 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8701 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8761 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8821 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8881 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     8941 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     9001 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     9061 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     9121 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     9181 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     9241 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     9301 tgccaagctg
//
