LOCUS       V029431                 7815 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029431
VERSION     V029431
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1009..2562
                     /label="Marco(NM_010766)"
                     /note="Marco(NM_010766)"
                     /gene="Marco"
     polyA_signal    2574..3196
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3238..3535
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3589..4383
                     /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    5035..5169
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(5175..5193)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(5207..5223)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5231..5247
                     /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(5255..5285)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5300..5321
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5609..6197)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6368..7228)
                     /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(7229..7333)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7360..7788
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgat gggaagtaaa
     1021 gaactcctca aagaggaaga cttcttgggc agcacagaag acagagccga ttttgaccaa
     1081 gctatgttcc ctgtgatgga gaccttcgaa atcaatgatc cagtgcccaa gaagagaaat
     1141 ggggggacct tctgcatggc agtcatggcc atccacctga tcctgctcac ggcaggtact
     1201 gcactgctgc tgattcaagt tctcaatctg caggagcagc tccagatgct agagatgtgc
     1261 tgtggcaatg gatcactagc tatcgaggac aagcccttct tctcgctgca gtgggcaccc
     1321 aaaacacacc tggtacctag agcacagggg ctgcaagcct tgcaggccca gctcagctgg
     1381 gtccatacca gccaggagca actccgtcag cagttcaaca acctcactca aaatccagag
     1441 ttgttccaga ttaaaggtga acgaggctct ccaggtccaa aaggggcccc gggtgctcct
     1501 ggaatccccg ggctgcctgg gccagctgct gagaagggag aaaagggggc tgcaggtcgt
     1561 gatggaaccc caggtgtcca aggaccccag ggcccaccag gcagcaaggg agaggcaggc
     1621 ctccagggac ttacgggtgc accagggaag caaggagcaa ctggtgctcc aggacctcga
     1681 ggagagaagg gcagcaaagg tgacataggt ctcactggcc ccaaggggga acatggcacc
     1741 aagggagaca aaggggacct aggccttcca ggaaacaaag gggacatggg catgaaggga
     1801 gacacggggc ccatggggtc ccctggagct cagggaggta aaggtgatgc tggaaaacca
     1861 ggcctaccag gtttggctgg atctccagga gtcaaaggtg accaaggaaa acctggagtg
     1921 cagggtgttc caggccctca aggtgcacca ggactttcag gtgccaaggg tgagccagga
     1981 cgcactggtc ttcctgggcc agcaggaccc ccgggaattg ctgggaatcc agggattgca
     2041 ggtgtgaaag gaagcaaggg tgacacagga attcaaggac agaaaggcac aaaaggagaa
     2101 tcaggagtcc caggtcttgt aggcagaaag ggagacactg gaagccctgg gctggcaggt
     2161 cccaaaggag aacctggacg agtcggtcag aagggagacc cggggatgaa agggtcttct
     2221 ggccagcaag gacaaaaggg agaaaagggt caaaaaggcg aatctttcca acgcgtccgg
     2281 atcatgggtg gcaccaacag aggccgagct gaagtttact ataacaatga gtgggggaca
     2341 atttgtgatg atgattggga taataatgat gcgactgtct tctgtcgcat gctcggttac
     2401 tccagaggga gagcacttag cagctatgga ggtggctctg ggaacatctg gctggacaat
     2461 gtgaattgtc ggggcacaga gaacagtttg tgggactgca gtaagaactc ctggggcaat
     2521 cacaattgcg tacataatga agatgcgggt gtggaatgct cctgaaggat cccgggtggc
     2581 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     2641 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     2701 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     2761 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     2821 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     2881 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     2941 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     3001 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa
     3061 aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac
     3121 cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt
     3181 agatgcctgt tgaattgggt acgcggccag cttggctgtg gaatgtgtgt cagttagggt
     3241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3301 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3361 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3421 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3481 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggaa
     3541 ttgatcagct tgggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3601 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3661 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3721 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3781 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3841 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3901 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3961 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4021 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4081 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4141 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4201 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4261 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4321 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4381 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4441 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4501 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccgggc
     4561 tcgatcccct cgcgagttgg ttcagctgct gcctgaggct ggacgacctc gcggagttct
     4621 accggcagtg caaatccgtc ggcatccagg aaaccagcag cggctatccg cgcatccatg
     4681 cccccgaact gcaggagtgg ggaggcacga tggccgcttt ggtcgacccg gacgggacgc
     4741 tcctgcgcct gatacagaac gaattgcttg caggcatctc atgagtgtgt cttcccgttt
     4801 tccgcctgag gtcactgcgt ggatggagcg ctggcgcctg ctgcgcgacg gcgagctgct
     4861 caccacccac tcgccaagct ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4921 gctccgccga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     4981 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     5041 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5101 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5161 gtctggatca attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc
     5221 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5281 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5341 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5401 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5461 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5521 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5581 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5641 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5701 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5761 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5821 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5881 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5941 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6001 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6061 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6121 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6181 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6241 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6301 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6361 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6421 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6481 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6541 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6601 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6661 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6721 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6781 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6841 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6901 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6961 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7021 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7081 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7141 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7201 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7261 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7321 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     7381 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     7441 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     7501 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     7561 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7621 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7681 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     7741 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     7801 taacgcttac aattt
//
