LOCUS       62056_22040        4985 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 4985)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        299..327
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             374..1081
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             1493..2284
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(2478..2511)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(2589..2605)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3079..3183
                     /label=AmpR promoter
     CDS             3184..4041
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4215..4803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 gccgcaactc cctgaaacgc ttccaggaac tgacgaagcg ccttctcagg cgagttaatt
      301 gacaattaat catcggctcg tataatgtgt ggaatgtctc atgagcggat atttcacaca
      361 ggaaacagta ttcatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
      421 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
      481 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      541 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
      601 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
      661 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
      721 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
      781 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
      841 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
      901 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
      961 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     1021 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     1081 gtaagaagtt cctatacttt ctagagaata ggaacttcgg aataggaact tcaagatccc
     1141 ctcacgctgc cgcaagcact cagggcgcaa gggctgctaa aggaagcgga acacgtagaa
     1201 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     1261 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     1321 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     1381 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     1441 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     1501 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1561 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1621 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     1681 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1741 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     1801 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     1861 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     1921 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     1981 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     2041 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2101 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     2161 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     2221 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     2281 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     2341 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     2401 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     2461 agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga
     2521 actaaaatta aggcggcagc aatgccgcct tttctttttc cgaaaactcc gtttctgcac
     2581 tagaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2641 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2701 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     2761 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     2821 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     2881 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     2941 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     3001 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3061 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3121 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3181 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3241 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3301 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3361 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3421 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3481 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3541 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3601 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3661 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3721 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3781 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3841 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3901 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3961 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4021 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4081 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4141 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4201 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4261 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4321 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4381 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4441 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4501 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4561 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4621 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4681 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4741 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4801 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4861 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4921 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4981 gaaga
//
