LOCUS       V041515                 9261 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041515
VERSION     V041515
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3313
                     /label="ABHD13(NM_032859)"
                     /note="ABHD13(NM_032859)"
                     /gene="ABHD13"
     CDS             3320..3409
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3459..3670
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3683..3951
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3986..4693
                     /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             4703..4756
                     /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&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4757..5356
                     /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    5357..5944
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6018..6251
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6323..6444
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6463..6598
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6636..6652)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6660..6676
                     /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(6684..6714)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6729..6750
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7038..7626)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7797..8657)
                     /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(8658..8762)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9236..9252
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatggaaaa gtcctggatg ctgtggaact ttgttgaaag
     2341 atggctaata gccttggctt catggtcttg ggctctctgc cgtatttctc ttttaccttt
     2401 aatagtgact tttcatctgt atggaggcat tatcttactt ttgttaatat tcatatcaat
     2461 agcaggtatt ctgtataaat tccaggatgt attgctttat tttccagaac agccatcctc
     2521 ttcacgtctt tatgttccca tgcccactgg cattccacat gaaaacattt tcatcagaac
     2581 caaagatgga atacgtctga atcttatttt gatacgatac actggagaca attcacccta
     2641 ttccccaact ataatttatt ttcatgggaa tgcaggcaac ataggtcaca ggttgccaaa
     2701 tgcattactt atgttggtta acctcaaagt taaccttttg ctggttgatt atcgaggata
     2761 tggaaaaagt gaaggagaag caagtgaaga aggactctac ttagattctg aagctgtgtt
     2821 agactacgtg atgactagac ctgaccttga taaaacaaaa atttttcttt ttggccgttc
     2881 cttgggtgga gcagtggcta ttcatttggc ttctgaaaat tcacatagga tttcagccat
     2941 tatggtggag aacacatttt taagcatacc acatatggcc agcactttat tttcattctt
     3001 tccgatgcgt taccttcctt tatggtgcta caaaaataaa tttttgtcct acagaaaaat
     3061 ctctcagtgt agaatgcctt cacttttcat ctctggactc tcagatcaat taattccacc
     3121 agtaatgatg aaacaacttt atgaactctc cccatctcgg actaagagat tagccatttt
     3181 tccagatggg actcacaatg acacatggca gtgccaaggc tatttcactg cacttgaaca
     3241 gttcatcaaa gaagtcgtaa agagccattc tcctgaagaa atggcaaaaa cttcatctaa
     3301 tgtaacaatt atactcgagg aacaaaaact aatatcagag gaagatttgg aacagaaact
     3361 aatcagtgaa gaagatttag agcaaaagtt aatttccgag gaggacttat aggatccgcg
     3421 gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     3481 agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg
     3541 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     3601 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     3661 cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac
     3721 ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc
     3781 ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc
     3841 ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct
     3901 tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg
     3961 accggcgcct acgctagacg ccaccatggt gagcaagggc gaggaggata acatggccat
     4021 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     4081 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     4141 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     4201 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     4261 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     4321 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     4381 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     4441 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     4501 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     4561 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     4621 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     4681 cgagctgtac aagggatctc gcgagggcag aggaagtctt ctaacatgcg gtgacgtgga
     4741 ggagaatccc ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     4801 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4861 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4921 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4981 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5041 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5101 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5161 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5221 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5281 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5341 caagcccggt gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5401 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5461 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5521 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5581 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     5641 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     5701 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     5761 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     5821 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     5881 gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctcccttt ggccgcctcc
     5941 ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     6001 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt
     6061 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     6121 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     6181 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     6241 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     6301 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     6361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6421 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     6481 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6541 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6601 gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6661 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6721 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6781 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6841 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6901 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6961 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7021 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7081 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7141 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7201 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7261 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7321 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7381 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7441 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7501 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7561 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7621 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7681 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7741 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7801 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7861 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7921 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7981 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8041 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8101 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8161 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8221 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8281 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8341 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8401 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8461 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8521 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8581 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8641 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8701 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8761 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     8821 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     8881 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     8941 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     9001 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     9061 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     9121 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     9181 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     9241 acgacggcca gtgccaagct g
//
