LOCUS       V040769                 6454 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040769
VERSION     V040769
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             928..1890
                     /label="MRGPRD(NM_198923)"
                     /note="MRGPRD(NM_198923)"
                     /gene="MRGPRD"
     CDS             1897..1920
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1927..2016
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     polyA_signal    2054..2278
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2324..2752
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2766..3095
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3162..3956
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4130..4251
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4300..4316)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4324..4340
                     /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(4348..4378)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4393..4414
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4702..5287)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5458..6318)
                     /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(6319..6423)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagctgaatt cgccaccatg aaccagactt tgaatagcag tgggaccgtg
      961 gagtcagccc taaactattc cagagggagc acagtgcaca cggcctacct ggtgctgagc
     1021 tccctggcca tgttcacctg cctgtgcggg atggcaggca acagcatggt gatctggctg
     1081 ctgggctttc gaatgcacag gaaccccttc tgcatctata tcctcaacct ggcggcagcc
     1141 gacctcctct tcctcttcag catggcttcc acgctcagcc tggaaaccca gcccctggtc
     1201 aataccactg acaaggtcca cgagctgatg aagagactga tgtactttgc ctacacagtg
     1261 ggcctgagcc tgctgacggc catcagcacc cagcgctgtc tctctgtcct cttccctatc
     1321 tggttcaagt gtcaccggcc caggcacctg tcagcctggg tgtgtggcct gctgtggaca
     1381 ctctgtctcc tgatgaacgg gttgacctct tccttctgca gcaagttctt gaaattcaat
     1441 gaagatcggt gcttcagggt ggacatggtc caggccgccc tcatcatggg ggtcttaacc
     1501 ccagtgatga ctctgtccag cctgaccctc tttgtctggg tgcggaggag ctcccagcag
     1561 tggcggcggc agcccacacg gctgttcgtg gtggtcctgg cctctgtcct ggtgttcctc
     1621 atctgttccc tgcctctgag catctactgg tttgtgctct actggttgag cctgccgccc
     1681 gagatgcagg tcctgtgctt cagcttgtca cgcctctcct cgtccgtaag cagcagcgcc
     1741 aaccccgtca tctacttcct ggtgggcagc cggaggagcc acaggctgcc caccaggtcc
     1801 ctggggactg tgctccaaca ggcgcttcgc gaggagcccg agctggaagg tggggagacg
     1861 cccaccgtgg gcaccaatga gatgggggct ctcgaggatt acaaggatga cgacgataag
     1921 ggatccagcg cctggtccca ccctcagttt gagaagggcg gaggctctgg cggcggaagc
     1981 ggaggatctg cttggagcca cccccagttc gaaaagtgat gagggcccgt ttaaacccgc
     2041 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2101 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2161 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2221 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2281 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2341 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2401 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2461 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2521 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2581 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2641 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2701 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     2761 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2821 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2881 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2941 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3001 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3061 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3121 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3181 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3241 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3301 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3361 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3421 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3481 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3541 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3601 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3661 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3721 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3781 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3841 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3901 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3961 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4021 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4081 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4141 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4201 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4261 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4321 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4381 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4441 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4501 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4561 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4621 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4681 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4861 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5101 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5221 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5281 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5341 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5401 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5461 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5521 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5581 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5641 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5701 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5761 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5821 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5881 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5941 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6001 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6061 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6121 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6181 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6241 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6301 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6361 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6421 gcgcacattt ccccgaaaag tgccacctga cgtc
//
