LOCUS       sequence-217134-        6617 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 6617)
  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 6617)
  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-BC
FEATURES             Location/Qualifiers
     source          1..6617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     558..577
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     805..823
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(827..870)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    873..894
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        895..929
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     RBS             2038..2049
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             4749..4760
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    5442..5462
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5463..5520
                     /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(5544..5561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5715..6303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5795..5814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6486..6580)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(6601..6617,1..643))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
       61 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
      121 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
      181 tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag
      241 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
      301 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
      361 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
      421 tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
      481 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
      541 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
      601 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
      661 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
      721 agttggaacc tcttacgtgc ccgatcaact cgagtgccac ctgacgtcta agaaaccatt
      781 attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa
      841 aaatccttag ctttcgctaa ggatgatttc tggaattcgc ggccgcttct agagtttacg
      901 gctagctcag tcctaggtac aatgctagct actagagtca cacaggaaag tatactagat
      961 gaaagaacag ctcaagccgc tgctagaaga caagacttac cttcgccact tctggcatcc
     1021 cgtgtgtacc cttaatgagt tcgaacgcgc caacgccagt gggcacggcc ccatgggcgt
     1081 caccttgcta ggcgagaaat tggtgttggc caggttaaat tcaaagatca ttgcggctgc
     1141 tgaccgatgt gctcatcgat cggcacagct ctccatcggc cgcgtttgca gcaacgccgg
     1201 aaaggactat ctcgaatgcc cgtatcacgg ctggcgctac gatgaggctg gggcctgtca
     1261 actgatccct gcttgccctg acaaaagcat ctctcctcgg gccaagattt cctcattcga
     1321 ttgtgaggtg aaatacgaca tcgtgtgggt acggctggat aacagcttcg attgcactca
     1381 gattccatac ctcagcgatt tcgataatcc cgacatgcag gtaatcgttg ccgattcgta
     1441 tatttgggag actgttgccg agcggcggtg ggagaacttt acagattttt cgcactttgc
     1501 cttcgtacac ccagggacgc tctatgatcc gtttttcgct agccacccaa ctgtttacgt
     1561 gaatcgcgtt gatggtgagt tgcaattcaa actcgctccg ccgcgtgaaa tgaaaggcat
     1621 cccgccagaa gcaccgatgg gtgacttcac ctaccgctgc acaatgccgt attcagtaaa
     1681 tcttgaaatc aaattgtgga aagatgactc acgtttcgtt ctttggacta ccgctagccc
     1741 tgtcgataac aagtcttgcc ggaattttat gattattgtg cgtgagaagg ataatcaacc
     1801 tgatcatatg cacctggctt tccagaagcg ggtgcttgac gaagaccagc ctgttatcga
     1861 atcgcaatgg cctctcgaaa tacagacctc ggaagtctcc gttgcaaccg ataaaatttc
     1921 cgtccagttc cgcaaatggc ataaagagct atctctgtca gccgttgaag gacgggaggc
     1981 gttccgcgat tccgtcttga cgaatgttat tgaagaagaa cagtaatact agagattaaa
     2041 gaggagaaat actagatgtc tactgaccaa gtaattttta acgactggca tccagtcgct
     2101 gctttggaag atgtatctct cgataaacga taccgctgtc gactactagg tcgtacagtt
     2161 agttatgtga aaacatctga tgctgttaat gctcattggg aagaaagtgc agatgaaatt
     2221 aagaccatcc gcgctaaaga aatctatggt cttctgtggc tctcctttgc cgacaaaccc
     2281 agtgagatgt tcgatattgc agagttcaag gaacctgatc gccgaatcgt cagcgctgga
     2341 tctgtgcgcg taaatgtctc aggactgcgt gctatcgaaa actttctgga catggctcat
     2401 ttcccttttg ttcatacgga tattttgggt gcagagccac tgacggaagt tgagccgtat
     2461 aacgtcaact atgatgaaac tgttgatgag atcttcgcta ctgagtgtaa gttcccgcaa
     2521 cctaaaggct cagctactgc tgtcgagcca attgatatgc agtatatata ccgcattaca
     2581 cggccatatt ccgccatttt atataaaact tgcccgcccg aaccgcatag atgggatgcc
     2641 ctcggtctgt ttattcaacc tgtagacgaa gattggtgca tagcgcatac aattatgtgc
     2701 tatgtggatg acgttaactc agatcaacaa cttcgccatt tccagcagac gatttttggc
     2761 caggacttga tgattcttat caaccaagtt ccaaagcgtc ttcccctggc tgcaagtcga
     2821 gagagcccag tccgggccga tgtgcttgca acagcttatc gtcgctggct gcgtgagaaa
     2881 ggtgtgcaat atggtgctct gcgggactaa tactagagtc acacaggaaa gtactagatg
     2941 aacaaacttg acgtcaacca gtggtttcct attgctacca ctgaagatct cccgaagcgc
     3001 catgtctttc atgccacgtt gttggggcaa gaaatggcca tctggcgcga tgactctggt
     3061 tcagttaatg cttgggagaa ccgctgcccg catagaggat tgcggttgac actgggtgct
     3121 aataccggta acgagttgcg gtgtcagtat catggatgga cttatgaaag cgggactggt
     3181 ggctgcactt ttgtcccagc ccatcgcgat gcaccacccc caaatgccgc gcgggttaat
     3241 acttttcctg tccgcgaaaa gcacggcttt atctggacga cattaggtca gccgccagga
     3301 gagcccattt caatcctcga tgacgctcag cttgtaaacg ctgtaaaaac aaatctgcat
     3361 agcgtagtta tagatgctga tattgacgga gttgtcagcg tcctacgtca gaatctttca
     3421 gcgttcatcg atgtgtttgg tgcggccagc gctgaagatc tgcatttgaa atccatgctg
     3481 caagatcgag ggattctggt aacaagatca ggctctattg ctattcattt ttatatgcag
     3541 cgctcaacca ttagtaaatg cgttgtacat gcgcaagtac ttactccggg acgtccagga
     3601 tacgaacttc aaaagaacta ctcgtatgcc atgaacgtta tccgcagggc agcagaagct
     3661 gtagctaccg acttgattag cattacagat atcagcgatc agactatcga aaagcttgaa
     3721 gtcgttagag aaaacatgac taaggctcct ccaacccact atatctgcga agtggttacg
     3781 cgtactcaag agacaggtga tattaactca tactggctga agcctatcgg ctacccacta
     3841 ccagcattca gtccagggat gcacatcagc atcacaacgc cggagggtag cattcgacaa
     3901 tattccctcg tgaacgggcc tgacgagcgt gaatccttca tcatcggtgt gaagaaagag
     3961 attcagtccc gtggcggctc cagatcaatg cacgaagatg tgaaggttgg aacgcaacta
     4021 aaagttacac ttccgaggaa cggttttcca ctcgtccaaa ccagaaaaca cccgattctc
     4081 gtagcaggtg gcatcggtat caccccaatt ttgtgtatgg cacaggctct ggatcagcaa
     4141 ggttcatcgt atgaaataca ttattttgct cgtgcatttg agcatgttcc attccaggat
     4201 cgactgactg cgttgggcga tcgtttgaat gtgcatcttg gcctcggccc agacgagact
     4261 agagcaaaac ttgccgacat catggagatt cataacgccc aagacgtaga tgtttacact
     4321 tgcggcccgc aaccaatgat cgaaactgta tctgctgtcg ctcttgctca tggcatcgct
     4381 gaagagtcca tccgatttga atttttcagt aaaaagaacg atgttcccgt ttctgatgaa
     4441 gaatatgagg ttgagctcaa aaaaactggt caaatattca ctgtctcgcc tggctctacg
     4501 ttgttgcaag cttgtttgga caacgatgtt cgtatcgaag cttcttgtga gcagggtgta
     4561 tgcgggactt gtataactcc agtcgtatcc ggcgatctcg agcatcatga cacttacctt
     4621 tctaagaaag aaagggaaag cggtaagtgg atcatgccgt gtgtttcgcg ctgcaagtcc
     4681 aaaaaaatcg ttctcgatct gtgaggccgg gcatcaccca cggtatggac gagctgtata
     4741 aactagagaa agaggagaaa tactagatga tcacgcttta tgactatgaa ctctcgggga
     4801 attgctacaa ggtgcggatg ctcctttcca tcctgaagct accgtacacg atagagacag
     4861 tggaattttt cccgtccagg gaacataaat caccggcctt cctgcgcatc aatccactcg
     4921 gccagttgcc ggtattgcgc gatgatgatt tcgtgttgcg cgatgcccag gccatcctgg
     4981 tttacctggc gaacaaatac gacaccagtt tgacctggta tccgacacat gcgcccgcca
     5041 tcctcggcca ggcaaatatg tggctcgcgt ttgccgatgg tttgaccggc tcgatttccg
     5101 ctgcacgttt gcatgatttg tttttttacg atttcaatgc agagcaatgc cgtgcccgcg
     5161 cgcacgaact actgcgcata ctggacgaac acctgtgggc ggcggaacaa aatggacatc
     5221 aatggctatg tccggccgcc gcacccacca tcgcggatct tgcctgcttc ccgtatatcg
     5281 cattgtcgga tgaagccggc gtttcactac tcgactaccc ggcggtacga cgctggctgg
     5341 accgtgtcaa acgcatcccc ggattcaccg tcatgtcggg gatttttcca acctcagctt
     5401 ccttcgatcc ggcctcactt gcggaagaaa cctgtcaata atactagtag cggccgctgc
     5461 agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt tttctttaaa accgaaaaga
     5521 ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5581 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5641 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5701 gcgttgctgg cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5761 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5821 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5881 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5941 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6001 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6061 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6121 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6181 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6241 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6301 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6361 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6421 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagctcgagg
     6481 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     6541 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     6601 ttacgccccg ccctgcc
//
