LOCUS       62056_6980        7729 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7729)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        110..489
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        490..693
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        738..756
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2165..2191
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             2222..2929
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    3035..3259
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3305..3733
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3747..4076
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4125..5147
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5280..5413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5450..5466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5474..5490)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5498..5528)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5543..5564)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5852..6437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6611..7468)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7469..7573)
                     /label=AmpR promoter
ORIGIN
        1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
       61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      661 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      721 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagtgccgc
      781 cgccatggtg tgcttccgcc tcttcccggt tccgggctca gggctcgttc tggtctgcct
      841 agtcctggga gctgtgcggt cttatgcagg taactcgact atgggcagtg gatacaccat
      901 ttggatgccc gagaatccga gaccagggac accttgtgac atttttacca atagcagagg
      961 gaagagagca tccaacgggg gcagtggatc tggatccggt ggctcgagtt tggaacttaa
     1021 tttgacagat tcagaaaatg ccacttgcct ttatgcaaaa tggcagatga atttcacagt
     1081 tcgctatgaa actacaaata aaacttataa aactgtaacc atttcagacc atggcactgt
     1141 gacatataat ggaagcattt gtggggatga tcagaatggt cccaaaatag cagtgcagtt
     1201 cggacctggc ttttcctgga ttgcgaattt taccaaggca gcatctactt attcaattga
     1261 cagcgtctca ttttcctaca acactggtga taacacaaca tttcctgatg ctgaagataa
     1321 aggaattctt actgttgatg aacttttggc catcagaatt ccattgaatg acctttttag
     1381 atgcaatagt ttatcaactt tggaaaagaa tgatgttgtc caacactact gggatgttct
     1441 tgtacaagct tttgtccaaa atggcacagt gagcacaaat gagttcctgt gtgataaaga
     1501 caaaacttca acagtggcac ccaccataca caccactgtg ccatctccta ctacaacacc
     1561 tactccaaag gaaaaaccag aagctggaac ctattcagtt aataatggca atgatacttg
     1621 cctgctggct accatggggc tgcagctgaa catcactcag gataaggttg cttcagttat
     1681 taacatcaac cccaatacaa ctcactccac aggcagctgc cgttctcaca ctgctctact
     1741 tagactcaat agcagcacta ttaagtatct agactttgtc tttgctgtga aaaatgaaaa
     1801 ccgattttat ctgaaggaag tgaacatcag catgtatttg gttaatggct ccgttttcag
     1861 cattgcaaat aacaatctca gctactggga tgcccccctg ggaagttctt atatgtgcaa
     1921 caaagagcag actgtttcag tgtctggagc atttcagata aatacctttg atctaagggt
     1981 tcagcctttc aatgtgacac aaggaaagta ttctacagcc caagagtgtt cgctggatga
     2041 tgacaccatt ctaatcccaa ttatagttgg tgctggtctt tcaggcttga ttatcgttat
     2101 agtgattgct tacgtaattg gcagaagaaa aagttatgct ggatatcaga ctctgggaag
     2161 cggataccca tacgatgtgc cagattacgc tggtggaggc ggatcaggag gcggtggatc
     2221 tatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     2281 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2341 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     2401 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     2461 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     2521 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     2581 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     2641 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     2701 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     2761 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     2821 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     2881 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt agtaaggatc
     2941 cactagtcca gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagtct
     3001 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3061 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3121 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3181 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3241 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3301 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3361 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3421 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3481 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3541 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3601 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3661 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3721 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3901 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3961 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4021 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4081 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgatgaaa aagcctgaac
     4141 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     4201 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     4261 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     4321 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     4381 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     4441 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     4501 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     4561 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     4621 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     4681 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     4741 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     4801 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     4861 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     4921 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     4981 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     5041 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     5101 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtgctac
     5161 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5221 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5281 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5341 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5401 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5461 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5521 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5581 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5641 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5701 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5761 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5821 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5881 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5941 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6001 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6061 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6121 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6181 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6241 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6301 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6361 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     6421 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6481 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6541 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6601 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6661 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6721 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6781 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6841 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6901 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6961 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7021 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7081 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7141 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7201 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7261 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7321 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7381 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7441 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7501 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7561 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7621 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7681 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagc
//
