LOCUS       sequence-214564-        5520 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 5520)
  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 5520)
  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-C
FEATURES             Location/Qualifiers
     source          1..5520
                     /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             47..58
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             2758..2769
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    3451..3471
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3472..3529
                     /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(3553..3570)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3724..4312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3804..3823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4495..4589)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4610..5269)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5184..5203
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     5431..5449
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5453..5496)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5499..5520
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tttacggcta gctcagtcct aggtacaatg ctagctacta gagattaaag aggagaaata
       61 ctagatgtct actgaccaag taatttttaa cgactggcat ccagtcgctg ctttggaaga
      121 tgtatctctc gataaacgat accgctgtcg actactaggt cgtacagtta gttatgtgaa
      181 aacatctgat gctgttaatg ctcattggga agaaagtgca gatgaaatta agaccatccg
      241 cgctaaagaa atctatggtc ttctgtggct ctcctttgcc gacaaaccca gtgagatgtt
      301 cgatattgca gagttcaagg aacctgatcg ccgaatcgtc agcgctggat ctgtgcgcgt
      361 aaatgtctca ggactgcgtg ctatcgaaaa ctttctggac atggctcatt tcccttttgt
      421 tcatacggat attttgggtg cagagccact gacggaagtt gagccgtata acgtcaacta
      481 tgatgaaact gttgatgaga tcttcgctac tgagtgtaag ttcccgcaac ctaaaggctc
      541 agctactgct gtcgagccaa ttgatatgca gtatatatac cgcattacac ggccatattc
      601 cgccatttta tataaaactt gcccgcccga accgcataga tgggatgccc tcggtctgtt
      661 tattcaacct gtagacgaag attggtgcat agcgcataca attatgtgct atgtggatga
      721 cgttaactca gatcaacaac ttcgccattt ccagcagacg atttttggcc aggacttgat
      781 gattcttatc aaccaagttc caaagcgtct tcccctggct gcaagtcgag agagcccagt
      841 ccgggccgat gtgcttgcaa cagcttatcg tcgctggctg cgtgagaaag gtgtgcaata
      901 tggtgctctg cgggactaat actagagtca cacaggaaag tactagatga acaaacttga
      961 cgtcaaccag tggtttccta ttgctaccac tgaagatctc ccgaagcgcc atgtctttca
     1021 tgccacgttg ttggggcaag aaatggccat ctggcgcgat gactctggtt cagttaatgc
     1081 ttgggagaac cgctgcccgc atagaggatt gcggttgaca ctgggtgcta ataccggtaa
     1141 cgagttgcgg tgtcagtatc atggatggac ttatgaaagc gggactggtg gctgcacttt
     1201 tgtcccagcc catcgcgatg caccaccccc aaatgccgcg cgggttaata cttttcctgt
     1261 ccgcgaaaag cacggcttta tctggacgac attaggtcag ccgccaggag agcccatttc
     1321 aatcctcgat gacgctcagc ttgtaaacgc tgtaaaaaca aatctgcata gcgtagttat
     1381 agatgctgat attgacggag ttgtcagcgt cctacgtcag aatctttcag cgttcatcga
     1441 tgtgtttggt gcggccagcg ctgaagatct gcatttgaaa tccatgctgc aagatcgagg
     1501 gattctggta acaagatcag gctctattgc tattcatttt tatatgcagc gctcaaccat
     1561 tagtaaatgc gttgtacatg cgcaagtact tactccggga cgtccaggat acgaacttca
     1621 aaagaactac tcgtatgcca tgaacgttat ccgcagggca gcagaagctg tagctaccga
     1681 cttgattagc attacagata tcagcgatca gactatcgaa aagcttgaag tcgttagaga
     1741 aaacatgact aaggctcctc caacccacta tatctgcgaa gtggttacgc gtactcaaga
     1801 gacaggtgat attaactcat actggctgaa gcctatcggc tacccactac cagcattcag
     1861 tccagggatg cacatcagca tcacaacgcc ggagggtagc attcgacaat attccctcgt
     1921 gaacgggcct gacgagcgtg aatccttcat catcggtgtg aagaaagaga ttcagtcccg
     1981 tggcggctcc agatcaatgc acgaagatgt gaaggttgga acgcaactaa aagttacact
     2041 tccgaggaac ggttttccac tcgtccaaac cagaaaacac ccgattctcg tagcaggtgg
     2101 catcggtatc accccaattt tgtgtatggc acaggctctg gatcagcaag gttcatcgta
     2161 tgaaatacat tattttgctc gtgcatttga gcatgttcca ttccaggatc gactgactgc
     2221 gttgggcgat cgtttgaatg tgcatcttgg cctcggccca gacgagacta gagcaaaact
     2281 tgccgacatc atggagattc ataacgccca agacgtagat gtttacactt gcggcccgca
     2341 accaatgatc gaaactgtat ctgctgtcgc tcttgctcat ggcatcgctg aagagtccat
     2401 ccgatttgaa tttttcagta aaaagaacga tgttcccgtt tctgatgaag aatatgaggt
     2461 tgagctcaaa aaaactggtc aaatattcac tgtctcgcct ggctctacgt tgttgcaagc
     2521 ttgtttggac aacgatgttc gtatcgaagc ttcttgtgag cagggtgtat gcgggacttg
     2581 tataactcca gtcgtatccg gcgatctcga gcatcatgac acttaccttt ctaagaaaga
     2641 aagggaaagc ggtaagtgga tcatgccgtg tgtttcgcgc tgcaagtcca aaaaaatcgt
     2701 tctcgatctg tgaggccggg catcacccac ggtatggacg agctgtataa actagagaaa
     2761 gaggagaaat actagatgat cacgctttat gactatgaac tctcggggaa ttgctacaag
     2821 gtgcggatgc tcctttccat cctgaagcta ccgtacacga tagagacagt ggaatttttc
     2881 ccgtccaggg aacataaatc accggccttc ctgcgcatca atccactcgg ccagttgccg
     2941 gtattgcgcg atgatgattt cgtgttgcgc gatgcccagg ccatcctggt ttacctggcg
     3001 aacaaatacg acaccagttt gacctggtat ccgacacatg cgcccgccat cctcggccag
     3061 gcaaatatgt ggctcgcgtt tgccgatggt ttgaccggct cgatttccgc tgcacgtttg
     3121 catgatttgt ttttttacga tttcaatgca gagcaatgcc gtgcccgcgc gcacgaacta
     3181 ctgcgcatac tggacgaaca cctgtgggcg gcggaacaaa atggacatca atggctatgt
     3241 ccggccgccg cacccaccat cgcggatctt gcctgcttcc cgtatatcgc attgtcggat
     3301 gaagccggcg tttcactact cgactacccg gcggtacgac gctggctgga ccgtgtcaaa
     3361 cgcatccccg gattcaccgt catgtcgggg atttttccaa cctcagcttc cttcgagccg
     3421 gcctcacttg cggaagaaac ctgtcaataa tactagtagc ggccgctgca gtccggcaaa
     3481 aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt
     3541 tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3601 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3661 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3721 gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3781 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3841 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3901 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3961 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4021 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4081 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4141 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4201 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4261 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4321 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4381 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4441 taaatcaatc taaagtatat atgagtaaac ttggtctgac agctcgaggc ttggattctc
     4501 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     4561 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
     4621 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     4681 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     4741 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     4801 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     4861 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     4921 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     4981 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     5041 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     5101 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     5161 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     5221 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     5281 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     5341 ggtgaaagtt ggaacctctt acgtgcccga tcaactcgag tgccacctga cgtctaagaa
     5401 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga
     5461 taaaaaaaat ccttagcttt cgctaaggat gatttctgga attcgcggcc gcttctagag
//
