LOCUS       sequence-217141-        6822 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 6822)
  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 6822)
  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-AB
FEATURES             Location/Qualifiers
     source          1..6822
                     /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             4060..4071
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    4753..4773
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4774..4831
                     /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(4855..4872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5026..5614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5106..5125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5797..5891)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5912..6571)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6486..6505
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     6733..6751
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6755..6798)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6801..6822
                     /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 atactagagt cacacaggaa agtactagat gaacaaactt gacgtcaacc agtggtttcc
     2281 tattgctacc actgaagatc tcccgaagcg ccatgtcttt catgccacgt tgttggggca
     2341 agaaatggcc atctggcgcg atgactctgg ttcagttaat gcttgggaga accgctgccc
     2401 gcatagagga ttgcggttga cactgggtgc taataccggt aacgagttgc ggtgtcagta
     2461 tcatggatgg acttatgaaa gcgggactgg tggctgcact tttgtcccag cccatcgcga
     2521 tgcaccaccc ccaaatgccg cgcgggttaa tacttttcct gtccgcgaaa agcacggctt
     2581 tatctggacg acattaggtc agccgccagg agagcccatt tcaatcctcg atgacgctca
     2641 gcttgtaaac gctgtaaaaa caaatctgca tagcgtagtt atagatgctg atattgacgg
     2701 agttgtcagc gtcctacgtc agaatctttc agcgttcatc gatgtgtttg gtgcggccag
     2761 cgctgaagat ctgcatttga aatccatgct gcaagatcga gggattctgg taacaagatc
     2821 aggctctatt gctattcatt tttatatgca gcgctcaacc attagtaaat gcgttgtaca
     2881 tgcgcaagta cttactccgg gacgtccagg atacgaactt caaaagaact actcgtatgc
     2941 catgaacgtt atccgcaggg cagcagaagc tgtagctacc gacttgatta gcattacaga
     3001 tatcagcgat cagactatcg aaaagcttga agtcgttaga gaaaacatga ctaaggctcc
     3061 tccaacccac tatatctgcg aagtggttac gcgtactcaa gagacaggtg atattaactc
     3121 atactggctg aagcctatcg gctacccact accagcattc agtccaggga tgcacatcag
     3181 catcacaacg ccggagggta gcattcgaca atattccctc gtgaacgggc ctgacgagcg
     3241 tgaatccttc atcatcggtg tgaagaaaga gattcagtcc cgtggcggct ccagatcaat
     3301 gcacgaagat gtgaaggttg gaacgcaact aaaagttaca cttccgagga acggttttcc
     3361 actcgtccaa accagaaaac acccgattct cgtagcaggt ggcatcggta tcaccccaat
     3421 tttgtgtatg gcacaggctc tggatcagca aggttcatcg tatgaaatac attattttgc
     3481 tcgtgcattt gagcatgttc cattccagga tcgactgact gcgttgggcg atcgtttgaa
     3541 tgtgcatctt ggcctcggcc cagacgagac tagagcaaaa cttgccgaca tcatggagat
     3601 tcataacgcc caagacgtag atgtttacac ttgcggcccg caaccaatga tcgaaactgt
     3661 atctgctgtc gctcttgctc atggcatcgc tgaagagtcc atccgatttg aatttttcag
     3721 taaaaagaac gatgttcccg tttctgatga agaatatgag gttgagctca aaaaaactgg
     3781 tcaaatattc actgtctcgc ctggctctac gttgttgcaa gcttgtttgg acaacgatgt
     3841 tcgtatcgaa gcttcttgtg agcagggtgt atgcgggact tgtataactc cagtcgtatc
     3901 cggcgatctc gagcatcatg acacttacct ttctaagaaa gaaagggaaa gcggtaagtg
     3961 gatcatgccg tgtgtttcgc gctgcaagtc caaaaaaatc gttctcgatc tgtgaggccg
     4021 ggcatcaccc acggtatgga cgagctgtat aaactagaga aagaggagaa atactagatg
     4081 atcacgcttt atgactatga actctcgggg aattgctaca aggtgcggat gctcctttcc
     4141 atcctgaagc taccgtacac gatagagaca gtggaatttt tcccgtccag ggaacataaa
     4201 tcaccggcct tcctgcgcat caatccactc ggccagttgc cggtattgcg cgatgatgat
     4261 ttcgtgttgc gcgatgccca ggccatcctg gtttacctgg cgaacaaata cgacaccagt
     4321 ttgacctggt atccgacaca tgcgcccgcc atcctcggcc aggcaaatat gtggctcgcg
     4381 tttgccgatg gtttgaccgg ctcgatttcc gctgcacgtt tgcatgattt gtttttttac
     4441 gatttcaatg cagagcaatg ccgtgcccgc gcgcacgaac tactgcgcat actggacgaa
     4501 cacctgtggg cggcggaaca aaatggacat caatggctat gtccggccgc cgcacccacc
     4561 atcgcggatc ttgcctgctt cccgtatatc gcattgtcgg atgaagccgg cgtttcacta
     4621 ctcgactacc cggcggtacg acgctggctg gaccgtgtca aacgcatccc cggattcacc
     4681 gtcatgtcgg ggatttttcc aacctcagct tccttcgatc cggcctcact tgcggaagaa
     4741 acctgtcaat aatactagta gcggccgctg cagtccggca aaaaagggca aggtgtcacc
     4801 accctgccct ttttctttaa aaccgaaaag attacttcgc gttatgcagg cttcctcgct
     4861 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4921 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4981 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg
     5041 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5101 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5161 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5221 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5281 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5341 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5401 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5461 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5521 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5581 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5641 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5701 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5761 atatgagtaa acttggtctg acagctcgag gcttggattc tcaccaataa aaaacgcccg
     5821 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
     5881 caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt
     5941 actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa
     6001 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga
     6061 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     6121 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     6181 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     6241 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     6301 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg
     6361 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     6421 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     6481 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     6541 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     6601 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     6661 ttacgtgccc gatcaactcg agtgccacct gacgtctaag aaaccattat tatcatgaca
     6721 ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct
     6781 ttcgctaagg atgatttctg gaattcgcgg ccgcttctag ag
//
