LOCUS       sequence-217137-        7706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Demethylates methylxanthines..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7706)
  AUTHORS   Gutierrez AE, Shah P, Rex AE, Nguyen TC, Kenkare SP, Barrick JE, 
            Mishler DM
  TITLE     Bioassay for Determining the Concentrations of Caffeine and 
            Individual Methylxanthines in Complex Samples
  JOURNAL   Appl Environ Microbiol. 2019 Nov 14;85(23). pii: AEM.01965-19. doi: 
            10.1128/AEM.01965-19. Print 2019 Dec 1.
  PUBMED    31540989
REFERENCE   2  (bases 1 to 7706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol."; date: "2019-11-14"; volume: "85(23). pii: 
            AEM.01965-19. doi"; pages: " 10.1128/AEM.01965-19. Print 2019 Dec 1"
COMMENT     Sequence Label: pDCAF-ABC
FEATURES             Location/Qualifiers
     source          1..7706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     RBS             53..61
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     RBS             2233..2244
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             4944..4955
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    5637..5657
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5658..5715
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(5739..5756)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5910..6498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5990..6009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6681..6775)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6796..7455)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7370..7389
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     7617..7635
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7639..7682)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7685..7706
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tttacggcta gctcagtcct aggtacaatg ctagctacta gagaaatcaa attaaggagg
       61 taagataaat ggaacaggca atcattaatg atgaacggga gtatcttcgc catttctggc
      121 atcccgtatg tactgtaact gagcttgaaa aggcgcatcc ttccagcctc ggccccctgg
      181 ccgttaagct gctgaatgaa cagctcgttg tcgccaagct aggcgatgag tacgtcgcga
      241 tgcgtgatag atgcgctcat cgatcagcta agctttcctt gggtacagta agtggaaacc
      301 gcctacagtg cccctatcac ggatggcaat atgatacgca tggcgcttgc cagctcgtac
      361 cagcgtgccc caacagccca atacccaaca aggctaaagt tgatcgcttc gattgcgaag
      421 aacgctatgg attgatttgg attcgattgg actctagttt tgactgcact gaaattccct
      481 actttagtgc agccaacgat cctaggttgc gtattgttat acaagaacct tactggtggg
      541 atgcgactgc ggaacgtaga tgggaaaatt ttacagattt ttctcacttt gcattcattc
      601 acccaggcac gcttttcgat ccaaataatg ctgaacctcc aattgttccg atggatcgat
      661 tcaatggtca gtttcggttt gtctacgaca cgccagaaga tatggctgtc ccaaatcagg
      721 ctccaattgg ttcattttcg tatacttgca gcatgccgtt tgctattaac cttgaagtat
      781 ccaaatactc cagcagttcg ctgcatgtgt tattcaatgt gtcatgtccg gtagacagcc
      841 acaccacgaa aaactttctg atcttcgcta gggagcaatc ggacgactcg gattatctgc
      901 acattgcatt taatgatctc gtcttcgctg aagacaaacc agtaattgag tcccaatggc
      961 ctaaagacgc gccagcagat gaggtctcag tagtcgcaga taaggtatcg atacaatata
     1021 gaaaatggct gcgggaacta aaagaagctc ataaagaagg ttcacaagcc ttccgaagtg
     1081 ctttgttaga cccagtcatt gaaagcgata ggagctacat ataatactag agtcacacag
     1141 gaaagtatac tagatgaaag aacagctcaa gccgctgcta gaagacaaga cttaccttcg
     1201 ccacttctgg catcccgtgt gtacccttaa tgagttcgaa cgcgccaacg ccagtgggca
     1261 cggccccatg ggcgtcacct tgctaggcga gaaattggtg ttggccaggt taaattcaaa
     1321 gatcattgcg gctgctgacc gatgtgctca tcgatcggca cagctctcca tcggccgcgt
     1381 ttgcagcaac gccggaaagg actatctcga atgcccgtat cacggctggc gctacgatga
     1441 ggctggggcc tgtcaactga tccctgcttg ccctgacaaa agcatctctc ctcgggccaa
     1501 gatttcctca ttcgattgtg aggtgaaata cgacatcgtg tgggtacggc tggataacag
     1561 cttcgattgc actcagattc catacctcag cgatttcgat aatcccgaca tgcaggtaat
     1621 cgttgccgat tcgtatattt gggagactgt tgccgagcgg cggtgggaga actttacaga
     1681 tttttcgcac tttgccttcg tacacccagg gacgctctat gatccgtttt tcgctagcca
     1741 cccaactgtt tacgtgaatc gcgttgatgg tgagttgcaa ttcaaactcg ctccgccgcg
     1801 tgaaatgaaa ggcatcccgc cagaagcacc gatgggtgac ttcacctacc gctgcacaat
     1861 gccgtattca gtaaatcttg aaatcaaatt gtggaaagat gactcacgtt tcgttctttg
     1921 gactaccgct agccctgtcg ataacaagtc ttgccggaat tttatgatta ttgtgcgtga
     1981 gaaggataat caacctgatc atatgcacct ggctttccag aagcgggtgc ttgacgaaga
     2041 ccagcctgtt atcgaatcgc aatggcctct cgaaatacag acctcggaag tctccgttgc
     2101 aaccgataaa atttccgtcc agttccgcaa atggcataaa gagctatctc tgtcagccgt
     2161 tgaaggacgg gaggcgttcc gcgattccgt cttgacgaat gttattgaag aagaacagta
     2221 atactagaga ttaaagagga gaaatactag atgtctactg accaagtaat ttttaacgac
     2281 tggcatccag tcgctgcttt ggaagatgta tctctcgata aacgataccg ctgtcgacta
     2341 ctaggtcgta cagttagtta tgtgaaaaca tctgatgctg ttaatgctca ttgggaagaa
     2401 agtgcagatg aaattaagac catccgcgct aaagaaatct atggtcttct gtggctctcc
     2461 tttgccgaca aacccagtga gatgttcgat attgcagagt tcaaggaacc tgatcgccga
     2521 atcgtcagcg ctggatctgt gcgcgtaaat gtctcaggac tgcgtgctat cgaaaacttt
     2581 ctggacatgg ctcatttccc ttttgttcat acggatattt tgggtgcaga gccactgacg
     2641 gaagttgagc cgtataacgt caactatgat gaaactgttg atgagatctt cgctactgag
     2701 tgtaagttcc cgcaacctaa aggctcagct actgctgtcg agccaattga tatgcagtat
     2761 atataccgca ttacacggcc atattccgcc attttatata aaacttgccc gcccgaaccg
     2821 catagatggg atgccctcgg tctgtttatt caacctgtag acgaagattg gtgcatagcg
     2881 catacaatta tgtgctatgt ggatgacgtt aactcagatc aacaacttcg ccatttccag
     2941 cagacgattt ttggccagga cttgatgatt cttatcaacc aagttccaaa gcgtcttccc
     3001 ctggctgcaa gtcgagagag cccagtccgg gccgatgtgc ttgcaacagc ttatcgtcgc
     3061 tggctgcgtg agaaaggtgt gcaatatggt gctctgcggg actaatacta gagtcacaca
     3121 ggaaagtact agatgaacaa acttgacgtc aaccagtggt ttcctattgc taccactgaa
     3181 gatctcccga agcgccatgt ctttcatgcc acgttgttgg ggcaagaaat ggccatctgg
     3241 cgcgatgact ctggttcagt taatgcttgg gagaaccgct gcccgcatag aggattgcgg
     3301 ttgacactgg gtgctaatac cggtaacgag ttgcggtgtc agtatcatgg atggacttat
     3361 gaaagcggga ctggtggctg cacttttgtc ccagcccatc gcgatgcacc acccccaaat
     3421 gccgcgcggg ttaatacttt tcctgtccgc gaaaagcacg gctttatctg gacgacatta
     3481 ggtcagccgc caggagagcc catttcaatc ctcgatgacg ctcagcttgt aaacgctgta
     3541 aaaacaaatc tgcatagcgt agttatagat gctgatattg acggagttgt cagcgtccta
     3601 cgtcagaatc tttcagcgtt catcgatgtg tttggtgcgg ccagcgctga agatctgcat
     3661 ttgaaatcca tgctgcaaga tcgagggatt ctggtaacaa gatcaggctc tattgctatt
     3721 catttttata tgcagcgctc aaccattagt aaatgcgttg tacatgcgca agtacttact
     3781 ccgggacgtc caggatacga acttcaaaag aactactcgt atgccatgaa cgttatccgc
     3841 agggcagcag aagctgtagc taccgacttg attagcatta cagatatcag cgatcagact
     3901 atcgaaaagc ttgaagtcgt tagagaaaac atgactaagg ctcctccaac ccactatatc
     3961 tgcgaagtgg ttacgcgtac tcaagagaca ggtgatatta actcatactg gctgaagcct
     4021 atcggctacc cactaccagc attcagtcca gggatgcaca tcagcatcac aacgccggag
     4081 ggtagcattc gacaatattc cctcgtgaac gggcctgacg agcgtgaatc cttcatcatc
     4141 ggtgtgaaga aagagattca gtcccgtggc ggctccagat caatgcacga agatgtgaag
     4201 gttggaacgc aactaaaagt tacacttccg aggaacggtt ttccactcgt ccaaaccaga
     4261 aaacacccga ttctcgtagc aggtggcatc ggtatcaccc caattttgtg tatggcacag
     4321 gctctggatc agcaaggttc atcgtatgaa atacattatt ttgctcgtgc atttgagcat
     4381 gttccattcc aggatcgact gactgcgttg ggcgatcgtt tgaatgtgca tcttggcctc
     4441 ggcccagacg agactagagc aaaacttgcc gacatcatgg agattcataa cgcccaagac
     4501 gtagatgttt acacttgcgg cccgcaacca atgatcgaaa ctgtatctgc tgtcgctctt
     4561 gctcatggca tcgctgaaga gtccatccga tttgaatttt tcagtaaaaa gaacgatgtt
     4621 cccgtttctg atgaagaata tgaggttgag ctcaaaaaaa ctggtcaaat attcactgtc
     4681 tcgcctggct ctacgttgtt gcaagcttgt ttggacaacg atgttcgtat cgaagcttct
     4741 tgtgagcagg gtgtatgcgg gacttgtata actccagtcg tatccggcga tctcgagcat
     4801 catgacactt acctttctaa gaaagaaagg gaaagcggta agtggatcat gccgtgtgtt
     4861 tcgcgctgca agtccaaaaa aatcgttctc gatctgtgag gccgggcatc acccacggta
     4921 tggacgagct gtataaacta gagaaagagg agaaatacta gatgatcacg ctttatgact
     4981 atgaactctc ggggaattgc tacaaggtgc ggatgctcct ttccatcctg aagctaccgt
     5041 acacgataga gacagtggaa tttttcccgt ccagggaaca taaatcaccg gccttcctgc
     5101 gcatcaatcc actcggccag ttgccggtat tgcgcgatga tgatttcgtg ttgcgcgatg
     5161 cccaggccat cctggtttac ctggcgaaca aatacgacac cagtttgacc tggtatccga
     5221 cacatgcgcc cgccatcctc ggccaggcaa atatgtggct cgcgtttgcc gatggtttga
     5281 ccggctcgat ttccgctgca cgtttgcatg atttgttttt ttacgatttc aatgcagagc
     5341 aatgccgtgc ccgcgcgcac gaactactgc gcatactgga cgaacacctg tgggcggcgg
     5401 aacaaaatgg acatcaatgg ctatgtccgg ccgccgcacc caccatcgcg gatcttgcct
     5461 gcttcccgta tatcgcattg tcggatgaag ccggcgtttc actactcgac tacccggcgg
     5521 tacgacgctg gctggaccgt gtcaaacgca tccccggatt caccgtcatg tcggggattt
     5581 ttccaacctc agcttccttc gatccggcct cacttgcgga agaaacctgt caataatact
     5641 agtagcggcc gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct
     5701 ttaaaaccga aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc
     5761 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5821 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5881 aaccgtaaaa aggccgcgtt gctggcgttt ttccacaggc tccgcccccc tgacgagcat
     5941 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6001 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6061 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6121 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6181 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6241 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6301 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6361 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6421 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6481 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6601 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6661 tctgacagct cgaggcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt
     6721 ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc
     6781 gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     6841 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     6901 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
     6961 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     7021 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     7081 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     7141 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     7201 ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag
     7261 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     7321 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     7381 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     7441 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     7501 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gcccgatcaa
     7561 ctcgagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7621 ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt
     7681 tctggaattc gcggccgctt ctagag
//
