LOCUS       V035394                 9494 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035394
VERSION     V035394
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        131..334
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             417..1811
                     /label="PRKN(NM_004562)"
                     /note="PRKN(NM_004562)"
                     /gene="PRKN"
     CDS             1827..2537
                     /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"
     misc_feature    2577..2694
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2743..3253
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     CDS             3275..3874
                     /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"
     LTR             4002..4235
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    4307..4428
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      4468..4603
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(4624..4642)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(4652..4668)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      4810..5265
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5291..5395
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             5396..6256
                     /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"
     rep_origin      6427..7015
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    7303..7324
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        7339..7369
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7377..7393
                     /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"
     primer_bind     7401..7417
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        7438..7456
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        7484..7710
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             7711..7891
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    7938..8063
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    8556..8789
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             8974..9018
                     /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"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgctaccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac gccaccatga
      421 tagtgtttgt caggttcaac tccagccatg gtttcccagt ggaggtcgat tctgacacca
      481 gcatcttcca gctcaaggag gtggttgcta agcgacaggg ggttccggct gaccagttgc
      541 gtgtgatttt cgcagggaag gagctgagga atgactggac tgtgcagaat tgtgacctgg
      601 atcagcagag cattgttcac attgtgcaga gaccgtggag aaaaggtcaa gaaatgaatg
      661 caactggagg cgacgacccc agaaacgcgg cgggaggctg tgagcgggag ccccagagct
      721 tgactcgggt ggacctcagc agctcagtcc tcccaggaga ctctgtgggg ctggctgtca
      781 ttctgcacac tgacagcagg aaggactcac caccagctgg aagtccagca ggtagatcaa
      841 tctacaacag cttttatgtg tattgcaaag gcccctgtca aagagtgcag ccgggaaaac
      901 tcagggtaca gtgcagcacc tgcaggcagg caacgctcac cttgacccag ggtccatctt
      961 gctgggatga tgttttaatt ccaaaccgga tgagtggtga atgccaatcc ccacactgcc
     1021 ctgggactag tgcagaattt ttctttaaat gtggagcaca ccccacctct gacaaggaaa
     1081 catcagtagc tttgcacctg atcgcaacaa atagtcggaa catcacttgc attacgtgca
     1141 cagacgtcag gagccccgtc ctggttttcc agtgcaactc ccgccacgtg atttgcttag
     1201 actgtttcca cttatactgt gtgacaagac tcaatgatcg gcagtttgtt cacgaccctc
     1261 aacttggcta ctccctgcct tgtgtggctg gctgtcccaa ctccttgatt aaagagctcc
     1321 atcacttcag gattctggga gaagagcagt acaaccggta ccagcagtat ggtgcagagg
     1381 agtgtgtcct gcagatgggg ggcgtgttat gcccccgccc tggctgtgga gcggggctgc
     1441 tgccggagcc tgaccagagg aaagtcacct gcgaaggggg caatggcctg ggctgtgggt
     1501 ttgccttctg ccgggaatgt aaagaagcgt accatgaagg ggagtgcagt gccgtatttg
     1561 aagcctcagg aacaactact caggcctaca gagtcgatga aagagccgcc gagcaggctc
     1621 gttgggaagc agcctccaaa gaaaccatca agaaaaccac caagccctgt ccccgctgcc
     1681 atgtaccagt ggaaaaaaat ggaggctgca tgcacatgaa gtgtccgcag ccccagtgca
     1741 ggctcgagtg gtgctggaac tgtggctgcg agtggaaccg cgtctgcatg ggggaccact
     1801 ggttcgacgt ggaaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca
     1861 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     1921 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     1981 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     2041 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     2101 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     2161 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     2221 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     2281 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     2341 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     2401 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     2461 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     2521 acgagctgta caagtgaatt ctcgacctcg agacaaatgg cagtattcat ccacaatttt
     2581 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2641 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     2701 tattacaggg acagcagaga tccactttgg ccgcggctcg agggggttgg ggttgcgcct
     2761 tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa
     2821 acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg
     2881 gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg
     2941 ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac
     3001 tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc
     3061 tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt
     3121 gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc
     3181 attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga
     3241 cctctctccc cagggggatc caccggagct taccatgacc gagtacaagc ccacggtgcg
     3301 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     3361 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     3421 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     3481 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     3541 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     3601 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     3661 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     3721 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     3781 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     3841 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     3901 gaccgaaagg agcgcacgac cccatgcatc ggtaccttta agaccaatga cttacaaggc
     3961 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     4021 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     4081 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     4141 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     4201 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     4261 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     4321 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4381 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     4441 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4501 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4561 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     4621 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     4681 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     4741 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4801 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4861 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4921 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4981 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5041 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5101 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5161 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5221 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     5281 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5341 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5401 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5461 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5521 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5581 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5641 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5701 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5761 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5821 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5881 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5941 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6001 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6061 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6121 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6181 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6241 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6301 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6361 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6421 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6481 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6541 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6601 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6661 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6721 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6781 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6841 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6901 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6961 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7021 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7081 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7141 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     7201 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7261 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     7321 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     7381 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     7441 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
     7501 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
     7561 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
     7621 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
     7681 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     7741 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     7801 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     7861 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     7921 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     7981 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     8041 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     8101 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     8161 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     8221 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     8281 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     8341 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     8401 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     8461 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     8521 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     8581 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     8641 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     8701 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     8761 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     8821 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     8881 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     8941 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     9001 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     9061 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     9121 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     9181 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     9241 agatccattc gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg
     9301 gccgccccct tcaccgaggg cctatttccc atgattcctt catatttgca tatacgatac
     9361 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
     9421 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     9481 aaaatggact atca
//
