LOCUS       sequence-217139-        6609 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 6609)
  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 6609)
  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-AC
FEATURES             Location/Qualifiers
     source          1..6609
                     /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             1136..1147
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             3847..3858
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    4540..4560
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4561..4618
                     /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(4642..4659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4813..5401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4893..4912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5584..5678)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5699..6358)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6273..6292
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     6520..6538
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6542..6585)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6588..6609
                     /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 agattaaaga
     1141 ggagaaatac tagatgtcta ctgaccaagt aatttttaac gactggcatc cagtcgctgc
     1201 tttggaagat gtatctctcg ataaacgata ccgctgtcga ctactaggtc gtacagttag
     1261 ttatgtgaaa acatctgatg ctgttaatgc tcattgggaa gaaagtgcag atgaaattaa
     1321 gaccatccgc gctaaagaaa tctatggtct tctgtggctc tcctttgccg acaaacccag
     1381 tgagatgttc gatattgcag agttcaagga acctgatcgc cgaatcgtca gcgctggatc
     1441 tgtgcgcgta aatgtctcag gactgcgtgc tatcgaaaac tttctggaca tggctcattt
     1501 cccttttgtt catacggata ttttgggtgc agagccactg acggaagttg agccgtataa
     1561 cgtcaactat gatgaaactg ttgatgagat cttcgctact gagtgtaagt tcccgcaacc
     1621 taaaggctca gctactgctg tcgagccaat tgatatgcag tatatatacc gcattacacg
     1681 gccatattcc gccattttat ataaaacttg cccgcccgaa ccgcatagat gggatgccct
     1741 cggtctgttt attcaacctg tagacgaaga ttggtgcata gcgcatacaa ttatgtgcta
     1801 tgtggatgac gttaactcag atcaacaact tcgccatttc cagcagacga tttttggcca
     1861 ggacttgatg attcttatca accaagttcc aaagcgtctt cccctggctg caagtcgaga
     1921 gagcccagtc cgggccgatg tgcttgcaac agcttatcgt cgctggctgc gtgagaaagg
     1981 tgtgcaatat ggtgctctgc gggactaata ctagagtcac acaggaaagt actagatgaa
     2041 caaacttgac gtcaaccagt ggtttcctat tgctaccact gaagatctcc cgaagcgcca
     2101 tgtctttcat gccacgttgt tggggcaaga aatggccatc tggcgcgatg actctggttc
     2161 agttaatgct tgggagaacc gctgcccgca tagaggattg cggttgacac tgggtgctaa
     2221 taccggtaac gagttgcggt gtcagtatca tggatggact tatgaaagcg ggactggtgg
     2281 ctgcactttt gtcccagccc atcgcgatgc accaccccca aatgccgcgc gggttaatac
     2341 ttttcctgtc cgcgaaaagc acggctttat ctggacgaca ttaggtcagc cgccaggaga
     2401 gcccatttca atcctcgatg acgctcagct tgtaaacgct gtaaaaacaa atctgcatag
     2461 cgtagttata gatgctgata ttgacggagt tgtcagcgtc ctacgtcaga atctttcagc
     2521 gttcatcgat gtgtttggtg cggccagcgc tgaagatctg catttgaaat ccatgctgca
     2581 agatcgaggg attctggtaa caagatcagg ctctattgct attcattttt atatgcagcg
     2641 ctcaaccatt agtaaatgcg ttgtacatgc gcaagtactt actccgggac gtccaggata
     2701 cgaacttcaa aagaactact cgtatgccat gaacgttatc cgcagggcag cagaagctgt
     2761 agctaccgac ttgattagca ttacagatat cagcgatcag actatcgaaa agcttgaagt
     2821 cgttagagaa aacatgacta aggctcctcc aacccactat atctgcgaag tggttacgcg
     2881 tactcaagag acaggtgata ttaactcata ctggctgaag cctatcggct acccactacc
     2941 agcattcagt ccagggatgc acatcagcat cacaacgccg gagggtagca ttcgacaata
     3001 ttccctcgtg aacgggcctg acgagcgtga atccttcatc atcggtgtga agaaagagat
     3061 tcagtcccgt ggcggctcca gatcaatgca cgaagatgtg aaggttggaa cgcaactaaa
     3121 agttacactt ccgaggaacg gttttccact cgtccaaacc agaaaacacc cgattctcgt
     3181 agcaggtggc atcggtatca ccccaatttt gtgtatggca caggctctgg atcagcaagg
     3241 ttcatcgtat gaaatacatt attttgctcg tgcatttgag catgttccat tccaggatcg
     3301 actgactgcg ttgggcgatc gtttgaatgt gcatcttggc ctcggcccag acgagactag
     3361 agcaaaactt gccgacatca tggagattca taacgcccaa gacgtagatg tttacacttg
     3421 cggcccgcaa ccaatgatcg aaactgtatc tgctgtcgct cttgctcatg gcatcgctga
     3481 agagtccatc cgatttgaat ttttcagtaa aaagaacgat gttcccgttt ctgatgaaga
     3541 atatgaggtt gagctcaaaa aaactggtca aatattcact gtctcgcctg gctctacgtt
     3601 gttgcaagct tgtttggaca acgatgttcg tatcgaagct tcttgtgagc agggtgtatg
     3661 cgggacttgt ataactccag tcgtatccgg cgatctcgag catcatgaca cttacctttc
     3721 taagaaagaa agggaaagcg gtaagtggat catgccgtgt gtttcgcgct gcaagtccaa
     3781 aaaaatcgtt ctcgatctgt gaggccgggc atcacccacg gtatggacga gctgtataaa
     3841 ctagagaaag aggagaaata ctagatgatc acgctttatg actatgaact ctcggggaat
     3901 tgctacaagg tgcggatgct cctttccatc ctgaagctac cgtacacgat agagacagtg
     3961 gaatttttcc cgtccaggga acataaatca ccggccttcc tgcgcatcaa tccactcggc
     4021 cagttgccgg tattgcgcga tgatgatttc gtgttgcgcg atgcccaggc catcctggtt
     4081 tacctggcga acaaatacga caccagtttg acctggtatc cgacacatgc gcccgccatc
     4141 ctcggccagg caaatatgtg gctcgcgttt gccgatggtt tgaccggctc gatttccgct
     4201 gcacgtttgc atgatttgtt tttttacgat ttcaatgcag agcaatgccg tgcccgcgcg
     4261 cacgaactac tgcgcatact ggacgaacac ctgtgggcgg cggaacaaaa tggacatcaa
     4321 tggctatgtc cggccgccgc acccaccatc gcggatcttg cctgcttccc gtatatcgca
     4381 ttgtcggatg aagccggcgt ttcactactc gactacccgg cggtacgacg ctggctggac
     4441 cgtgtcaaac gcatccccgg attcaccgtc atgtcgggga tttttccaac ctcagcttcc
     4501 ttcgatccgg cctcacttgc ggaagaaacc tgtcaataat actagtagcg gccgctgcag
     4561 tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
     4621 acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4681 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4741 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4801 gttgctggcg tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4921 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4981 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5221 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5521 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gctcgaggct
     5581 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     5641 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     5701 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     5761 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     5821 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     5881 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     5941 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     6001 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     6061 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     6121 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
     6181 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     6241 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     6301 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     6361 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     6421 tttcattatg gtgaaagttg gaacctctta cgtgcccgat caactcgagt gccacctgac
     6481 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag
     6541 aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttcgcggccg
     6601 cttctagag
//
