LOCUS       V041572                 9811 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041572
VERSION     V041572
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..4440
                     /label="Capn1(NM_001110504)"
                     /note="Capn1(NM_001110504)"
                     /gene="Capn1"
     CDS             4447..4536
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4580..4791
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4804..5072
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5105..5899
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5906..6494
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6568..6801
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6873..6994
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7013..7148
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7186..7202)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7210..7226
                     /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(7234..7264)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7279..7300
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7588..8176)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8347..9207)
                     /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(9208..9312)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9786..9802
                     /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 catgacagag gagttaatca cccctgtgta ctgcaccggg
     2341 gtgtcagccc aagtacagaa gaagcgggac aaggagctgg gcttgggccg ccatgaaaac
     2401 gccatcaagt acctgggcca ggattatgaa acgcttcggg caagatgcct gcagagtggg
     2461 gtcctcttcc aagacgaggc cttccctccg gtttctcata gcctgggctt caaggaactg
     2521 ggtcctcatt cctctaaaac ctatggcatc aaatggaagc ggcctacgga actgatgtca
     2581 aacccccagt tcatcgtgga tggagccacc cggacagaca tctgccaggg agcactgggg
     2641 gactgttggc tcctggctgc cattgcctcc ctcaccctca acgagactat tctgcaccga
     2701 gtggttccct acggccagag ctttcaggat ggctacgctg gcatctttca tttccagctg
     2761 tggcagttcg gggagtgggt cgacgtggtc atagatgact tgctgcccac caaggacggg
     2821 aagctggtgt tcgtgcactc tgcccagggc aacgaattct ggagcgcact gctggagaaa
     2881 gcctatgcta aagtgaatgg cagctatgag gctctttcgg gaggctgcac ctcagaggcc
     2941 ttcgaggact ttaccggtgg ggtcactgag tggtacgacc tgcagaaggc ccccagcgac
     3001 ctctaccaga tcattctaaa ggccctggaa cgaggctcct tgctgggctg ctccattaat
     3061 atctccgata tccgtgattt agaggctatt acttttaaga acctggtgag gggccatgcg
     3121 tactctgtga cgggcgccaa gcaggtaact taccagggcc agcgggtgaa cctaatccgg
     3181 atgcggaacc cctggggtga agtggagtgg aaaggaccct ggagtgacag ctcctatgag
     3241 tggaacaaag tggaccccta tgaacgagag cagctgaggg tcaagatgga ggatggggag
     3301 ttctggatgt cgttccgaga cttcatccgt gaattcacca aactggaaat ctgcaacctt
     3361 acaccggacg cccttaagag caggaccctc cggaattgga ataccacatt ttacgagggc
     3421 acctggcgtc ggggaagcac cgctggaggc tgcaggaact acccagctac cttctgggta
     3481 aacccccagt tcaagatccg gttggaggag gtggatgacg cagacgacta tgacaaccgg
     3541 gagtcgggct gcagcttctt gctggccctc atgcagaaac accgccgcag ggagcgtcgc
     3601 tttggccggg acatggagac catcggtttt gcagtgtacc aggtccctcg ggagctggcg
     3661 ggtcagcctg tgcacttgaa gcgtgatttc ttcctggcca acgcttctcg ggcgcagtca
     3721 gagcacttca tcaaccttcg ggaagtcagc aaccgtatcc gcctgccgcc cggggagtac
     3781 atagtggtgc cctccacctt cgagcccaac aaagaaggcg acttcctgct gcgcttcttc
     3841 tcagagaaga aggctgggac ccaggaacta gatgaccaga tccaggccaa cctccctgat
     3901 gagaaagttc tctctgaaga ggagattgat gacaacttca agaccttgtt cagcaagttg
     3961 gcaggggatg acatggagat cagcgtcaag gagctacaga ccatcttgaa caggatcatt
     4021 agcaaacaca aagacctgcg cactaatggc ttcagcctgg agtcgtgccg cagcatggtg
     4081 aacctcatgg atcgagatgg caacgggaag ctgggtctgg tggagttcaa catcctgtgg
     4141 aaccgcatcc gaaattacct gaccatcttc cggaagtttg acctggacaa gtctggcagc
     4201 atgagtgcct atgagatgag gatggccatc gaggctgcag gcttcaagct taacaagaag
     4261 ctgcatgaac tcataatcac ccgctactcg gagcccgacc tggccgtgga ctttgacaac
     4321 tttgtgtgct gtcttgtgcg gctggagacc atgttccggt ttttcaaact tctggacaca
     4381 gacctggatg gtgttgtgac ctttgatcta tttaagtggc tccagctgac tatgtttgcc
     4441 ggatccgaac aaaaactaat atcagaggaa gatttggaac agaaactaat cagtgaagaa
     4501 gatttagagc aaaagttaat ttccgaggag gacttatagc ggccgcgaag gatctgcgat
     4561 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     4621 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     4681 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     4741 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt
     4801 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc
     4861 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc
     4921 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta
     4981 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt
     5041 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctaggc
     5101 caccatggct gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5161 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5221 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5281 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5341 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5401 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5461 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5521 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5581 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5641 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5701 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5761 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5821 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5881 tcttgacgag ttcttctgag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5941 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     6001 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     6061 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     6121 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     6181 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     6241 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     6301 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     6361 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6421 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     6481 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     6541 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     6601 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     6661 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6721 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6781 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6841 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6901 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6961 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     7021 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     7081 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     7141 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     7201 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7261 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7321 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7381 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7441 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7501 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7561 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7621 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7681 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7741 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7801 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7861 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7921 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7981 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     8041 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8101 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8161 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8221 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8281 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8341 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8401 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8461 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8521 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8581 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8641 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8701 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8761 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8821 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8881 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8941 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     9001 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9061 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9121 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9181 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9241 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9301 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     9361 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     9421 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     9481 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     9541 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     9601 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     9661 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9721 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9781 acgttgtaaa acgacggcca gtgccaagct g
//
