LOCUS       V031741                 6652 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031741
VERSION     V031741
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             917..1441
                     /label="ARF6(NM_001663)"
                     /note="ARF6(NM_001663)"
                     /gene="ARF6"
     CDS             1466..2176
                     /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"
     promoter        complement(2204..2222)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     polyA_signal    2248..2472
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2518..2946
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2960..3290
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3357..4151
                     /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    4325..4446
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4495..4511)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4519..4535
                     /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(4543..4573)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4588..4609
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4897..5485)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5656..6516)
                     /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(6517..6621)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat gccaccatgg ggaaggtgct atccaaaatc ttcgggaaca aggaaatgcg
      961 gatcctcatg ttgggcctgg acgcggccgg caagacaaca atcctgtaca agttgaagct
     1021 gggccagtcg gtgaccacca ttcccactgt gggtttcaac gtggagacgg tgacttacaa
     1081 aaatgtcaag ttcaacgtat gggatgtggg cggccaggac aagatccggc cgctctggcg
     1141 gcattactac actgggaccc aaggtctcat cttcgtagtg gactgcgccg accgcgaccg
     1201 catcgatgag gctcgccagg agctgcaccg cattatcaat gaccgggaga tgagggacgc
     1261 cataatcctc atcttcgcca acaagcagga cctgcccgat gccatgaaac cccacgagat
     1321 ccaggagaaa ctgggcctga cccggattcg ggacaggaac tggtatgtgc agccctcctg
     1381 tgccacctca ggggacggac tctatgaggg gctcacatgg ttaacctcta actacaaatc
     1441 tctcgaggga cttaggtctg ccaccatggt gagcaagggc gaggaggata acatggccat
     1501 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     1561 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     1621 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     1681 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     1741 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     1801 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     1861 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     1921 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     1981 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     2041 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     2101 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     2161 cgagctgtac aagtaactcg agcatgcatc tagagggccc tattctatag tgtcacctaa
     2221 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2281 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2341 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2401 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2461 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2521 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2581 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2641 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg
     2701 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2761 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2821 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2881 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2941 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg
     3001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3301 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3361 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3421 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3481 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3541 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3601 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3661 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3721 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3781 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3841 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3901 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3961 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4021 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4081 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4141 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4201 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4261 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4321 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4441 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4501 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4561 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4621 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4681 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     5101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5401 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6481 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
