LOCUS       sequence-214565-        5739 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 5739)
  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 5739)
  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-B
FEATURES             Location/Qualifiers
     source          1..5739
                     /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             2971..2982
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    3664..3684
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3691..3748
                     /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(3772..3789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3943..4531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4023..4042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4714..4808)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4829..5488)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5403..5422
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     5650..5668
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5672..5715)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5718..5739
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tttacggcta gctcagtcct aggtacaatg ctagctacta gagtcacaca ggaaagtata
       61 ctagatgaaa gaacagctca agccgctgct agaagacaag acttaccttc gccacttctg
      121 gcatcccgtg tgtaccctta atgagttcga acgcgccaac gccagtgggc acggccccat
      181 gggcgtcacc ttgctaggcg agaaattggt gttggccagg ttaaattcaa agatcattgc
      241 ggctgctgac cgatgtgctc atcgatcggc acagctctcc atcggccgcg tttgcagcaa
      301 cgccggaaag gactatctcg aatgcccgta tcacggctgg cgctacgatg aggctggggc
      361 ctgtcaactg atccctgctt gccctgacaa aagcatctct cctcgggcca agatttcctc
      421 attcgattgt gaggtgaaat acgacatcgt gtgggtacgg ctggataaca gcttcgattg
      481 cactcagatt ccatacctca gcgatttcga taatcccgac atgcaggtaa tcgttgccga
      541 ttcgtatatt tgggagactg ttgccgagcg gcggtgggag aactttacag atttttcgca
      601 ctttgccttc gtacacccag ggacgctcta tgatccgttt ttcgctagcc acccaactgt
      661 ttacgtgaat cgcgttgatg gtgagttgca attcaaactc gctccgccgc gtgaaatgaa
      721 aggcatcccg ccagaagcac cgatgggtga cttcacctac cgctgcacaa tgccgtattc
      781 agtaaatctt gaaatcaaat tgtggaaaga tgactcacgt ttcgttcttt ggactaccgc
      841 tagccctgtc gataacaagt cttgccggaa ttttatgatt attgtgcgtg agaaggataa
      901 tcaacctgat catatgcacc tggctttcca gaagcgggtg cttgacgaag accagcctgt
      961 tatcgaatcg caatggcctc tcgaaataca gacctcggaa gtctccgttg caaccgataa
     1021 aatttccgtc cagttccgca aatggcataa agagctatct ctgtcagccg ttgaaggacg
     1081 ggaggcgttc cgcgattccg tcttgacgaa tgttattgaa gaagaacagt aatactagag
     1141 tcacacagga aagtactaga tgaacaaact tgacgtcaac cagtggtttc ctattgctac
     1201 cactgaagat ctcccgaagc gccatgtctt tcatgccacg ttgttggggc aagaaatggc
     1261 catctggcgc gatgactctg gttcagttaa tgcttgggag aaccgctgcc cgcatagagg
     1321 attgcggttg acactgggtg ctaataccgg taacgagttg cggtgtcagt atcatggatg
     1381 gacttatgaa agcgggactg gtggctgcac ttttgtccca gcccatcgcg atgcaccacc
     1441 cccaaatgcc gcgcgggtta atacttttcc tgtccgcgaa aagcacggct ttatctggac
     1501 gacattaggt cagccgccag gagagcccat ttcaatcctc gatgacgctc agcttgtaaa
     1561 cgctgtaaaa acaaatctgc atagcgtagt tatagatgct gatattgacg gagttgtcag
     1621 cgtcctacgt cagaatcttt cagcgttcat cgatgtgttt ggtgcggcca gcgctgaaga
     1681 tctgcatttg aaatccatgc tgcaagatcg agggattctg gtaacaagat caggctctat
     1741 tgctattcat ttttatatgc agcgctcaac cattagtaaa tgcgttgtac atgcgcaagt
     1801 acttactccg ggacgtccag gatacgaact tcaaaagaac tactcgtatg ccatgaacgt
     1861 tatccgcagg gcagcagaag ctgtagctac cgacttgatt agcattacag atatcagcga
     1921 tcagactatc gaaaagcttg aagtcgttag agaaaacatg actaaggctc ctccaaccca
     1981 ctatatctgc gaagtggtta cgcgtactca agagacaggt gatattaact catactggct
     2041 gaagcctatc ggctacccac taccagcatt cagtccaggg atgcacatca gcatcacaac
     2101 gccggagggt agcattcgac aatattccct cgtgaacggg cctgacgagc gtgaatcctt
     2161 catcatcggt gtgaagaaag agattcagtc ccgtggcggc tccagatcaa tgcacgaaga
     2221 tgtgaaggtt ggaacgcaac taaaagttac acttccgagg aacggttttc cactcgtcca
     2281 aaccagaaaa cacccgattc tcgtagcagg tggcatcggt atcaccccaa ttttgtgtat
     2341 ggcacaggct ctggatcagc aaggttcatc gtatgaaata cattattttg ctcgtgcatt
     2401 tgagcatgtt ccattccagg atcgactgac tgcgttgggc gatcgtttga atgtgcatct
     2461 tggcctcggc ccagacgaga ctagagcaaa acttgccgac atcatggaga ttcataacgc
     2521 ccaagacgta gatgtttaca cttgcggccc gcaaccaatg atcgaaactg tatctgctgt
     2581 cgctcttgct catggcatcg ctgaagagtc catccgattt gaatttttca gtaaaaagaa
     2641 cgatgttccc gtttctgatg aagaatatga ggttgagctc aaaaaaactg gtcaaatatt
     2701 cactgtctcg cctggctcta cgttgttgca agcttgtttg gacaacgatg ttcgtatcga
     2761 agcttcttgt gagcagggtg tatgcgggac ttgtataact ccagtcgtat ccggcgatct
     2821 cgagcatcat gacacttacc tttctaagaa agaaagggaa agcggtaagt ggatcatgcc
     2881 gtgtgtttcg cgctgcaagt ccaaaaaaat cgttctcgat ctgtgaggcc gggcatcacc
     2941 cacggtatgg acgagctgta taaactagag aaagaggaga aatactagat gatcacgctt
     3001 tatgactatg aactctcggg gaattgctac aaggtgcgga tgctcctttc catcctgaag
     3061 ctaccgtaca cgatagagac agtggaattt ttcccgtcca gggaacataa atcaccggcc
     3121 ttcctgcgca tcaatccact cggccagttg ccggtattgc gcgatgatga tttcgtgttg
     3181 cgcgatgccc aggccatcct ggtttacctg gcgaacaaat acgacaccag tttgacctgg
     3241 tatccgacac atgcgcccgc catcctcggc caggcaaata tgtggctcgc gtttgccgat
     3301 ggtttgaccg gctcgatttc cgctgcacgt ttgcatgatt tgttttttta cgatttcaat
     3361 gcagagcaat gccgtgcccg cgcgcacgaa ctactgcgca tactggacga acacctgtgg
     3421 gcggcggaac aaaatggaca tcaatggcta tgtccggccg ccgcacccac catcgcggat
     3481 cttgcctgct tcccgtatat cgcattgtcg gatgaagccg gcgtttcact actcgactac
     3541 ccggcggtac gacgctggct ggaccgtgtc aaacgcatcc ccggattcac cgtcatgtcg
     3601 gggatttttc caacctcagc ttccttcgat ccggcctcac ttgcggaaga aacctgtcaa
     3661 taatactagt agcggccgct gcagatatgc tccggcaaaa aagggcaagg tgtcaccacc
     3721 ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac
     3781 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3841 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3901 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccaca ggctccgccc
     3961 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4021 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4081 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4141 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4201 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4261 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4321 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4381 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4441 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4501 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4561 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4621 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4681 tgagtaaact tggtctgaca gctcgaggct tggattctca ccaataaaaa acgcccggcg
     4741 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     4801 caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca tcgcagtact
     4861 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
     4921 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
     4981 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
     5041 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
     5101 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
     5161 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
     5221 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga aattccggat
     5281 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     5341 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     5401 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     5461 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     5521 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     5581 cgtgcccgat caactcgagt gccacctgac gtctaagaaa ccattattat catgacatta
     5641 acctataaaa ataggcgtat cacgaggcag aatttcagat aaaaaaaatc cttagctttc
     5701 gctaaggatg atttctggaa ttcgcggccg cttctagag
//
