LOCUS       V027039                 8024 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027039
VERSION     V027039
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             941..3514
                     /label="EEF2(NM_001961)"
                     /note="EEF2(NM_001961)"
                     /gene="EEF2"
     CDS             3521..3586
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3624..3848
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3894..4322
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4336..4665
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4732..5526
                     /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    5700..5821
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5870..5886)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5894..5910
                     /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(5918..5948)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5963..5984
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6272..6857)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7028..7888)
                     /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(7889..7993)
                     /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 aagcttggta ccgagctcgg atccgccacc atggtgaact tcacggtaga
      961 ccagatccgc gccatcatgg acaagaaggc caacatccgc aacatgtctg tcatcgccca
     1021 cgtggaccat ggcaagtcca cgctgacaga ctccctggtg tgcaaggcgg gcatcatcgc
     1081 ctcggcccgg gccggggaga cacgcttcac tgatacccgg aaggacgagc aggagcgttg
     1141 catcaccatc aagtcaactg ccatctccct cttctacgag ctctcggaga atgacttgaa
     1201 cttcatcaag cagagcaagg acggtgccgg cttcctcatc aacctcattg actcccccgg
     1261 gcatgtcgac ttctcctcgg aggtgactgc tgccctccga gtcaccgatg gcgcattggt
     1321 ggtggtggac tgcgtgtcag gcgtgtgcgt gcagacggag acagtgctgc ggcaggccat
     1381 tgccgagcgc atcaagcctg tgctgatgat gaacaagatg gaccgcgccc tgctggagct
     1441 gcagctggag cccgaggagc tctaccagac tttccagcgc atcgtggaga acgtgaacgt
     1501 catcatctcc acctacggcg agggcgagag cggccccatg ggcaacatca tgatcgatcc
     1561 tgtcctcggt accgtgggct ttgggtctgg cctccacggg tgggccttca ccctgaagca
     1621 gtttgccgag atgtatgtgg ccaagttcgc cgccaagggg gagggccagt tggggcctgc
     1681 cgagcgggcc aagaaagtag aggacatgat gaagaagctg tggggtgaca ggtactttga
     1741 cccagccaac ggcaagttca gcaagtcagc caccagcccc gaagggaaga agctgccacg
     1801 caccttctgc cagctgatcc tggaccccat cttcaaggtg tttgatgcga tcatgaattt
     1861 caagaaagag gagacagcaa aactgataga gaaactggac atcaaactgg acagcgagga
     1921 caaggacaaa gaaggcaaac ccctgctgaa ggctgtgatg cgccgctggc tgcctgccgg
     1981 agacgccttg ttgcagatga tcaccatcca cctgccctcc cctgtgacgg cccagaagta
     2041 ccgctgcgag ctcctgtacg aggggccccc ggacgacgag gctgccatgg gcattaaaag
     2101 ctgtgacccc aaaggccctc ttatgatgta tatttccaaa atggtgccaa cctccgacaa
     2161 aggtcggttc tacgcctttg gacgagtctt ctcggggctg gtctccactg gcctgaaggt
     2221 caggatcatg gggcccaact atacccctgg gaagaaggag gacctctacc tgaagccaat
     2281 ccagagaaca atcttgatga tgggccgcta cgtggagccc atcgaggatg tgccttgtgg
     2341 gaacattgtg ggcctcgtgg gcgtggacca gttcctggtg aagacgggca ccatcaccac
     2401 cttcgagcac gcgcacaaca tgcgggtgat gaagttcagc gtcagccctg ttgtcagagt
     2461 ggccgtggag gccaagaacc cggctgacct gcccaagctg gtggaggggc tgaagcggct
     2521 ggccaagtcc gaccccatgg tgcagtgcat catcgaggag tcgggagagc atatcatcgc
     2581 gggcgccggc gagctgcacc tggagatctg cctgaaggac ctggaggagg accacgcctg
     2641 catccccatc aagaaatctg acccggtcgt ctcgtaccgc gagacggtca gtgaagagtc
     2701 gaacgtgctc tgcctctcca agtcccccaa caagcacaac cggctgtaca tgaaggcgcg
     2761 gcccttcccc gacggcctgg ccgaggacat cgataaaggc gaggtgtccg cccgtcagga
     2821 gctcaagcag cgggcgcgct acctggccga gaagtacgag tgggacgtgg ctgaggcccg
     2881 caagatctgg tgctttgggc ccgacggcac cggccccaac atcctcaccg acatcaccaa
     2941 gggtgtgcag tacctcaacg agatcaagga cagtgtggtg gccggcttcc agtgggccac
     3001 caaggagggc gcactgtgtg aggagaacat gcggggtgtg cgcttcgacg tccacgacgt
     3061 caccctgcac gccgacgcca tccaccgcgg agggggccag atcatcccca cagcacggcg
     3121 ctgcctctat gccagtgtgc tgaccgccca gccacgcctc atggagccca tctaccttgt
     3181 ggagatccag tgtccagagc aggtggtcgg tggcatctac ggggttttga acaggaagcg
     3241 gggccacgtg ttcgaggagt cccaggtggc cggcaccccc atgtttgtgg tcaaggccta
     3301 tctgcccgtc aacgagtcct ttggcttcac cgctgacctg aggtccaaca cgggcggcca
     3361 ggcgttcccc cagtgtgtgt ttgaccactg gcagatcctg cccggagacc ccttcgacaa
     3421 cagcagccgc cccagccagg tggtggcgga gacccgcaag cgcaagggcc tgaaagaagg
     3481 catccctgcc ctggacaact tcctggacaa attgctcgag gactacaaag accatgacgg
     3541 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt
     3601 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3661 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3721 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3781 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3841 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3901 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3961 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4021 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4081 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4141 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4201 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4261 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4321 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4381 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4441 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4501 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4561 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4621 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4681 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4741 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4801 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4861 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4921 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4981 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5041 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5101 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5161 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5221 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5281 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5341 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5401 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5461 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5521 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5581 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5641 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5701 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5761 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5821 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5881 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5941 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6001 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6061 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6781 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     6841 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6901 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6961 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7021 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7081 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7141 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7201 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7261 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7321 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7381 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7441 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7501 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7561 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7621 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7681 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7741 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7801 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7861 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7921 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7981 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
