LOCUS       sequence-214562-        5725 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 5725)
  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 5725)
  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-A
FEATURES             Location/Qualifiers
     source          1..5725
                     /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             2963..2974
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    3656..3676
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3677..3734
                     /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(3758..3775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3929..4517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4009..4028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4700..4794)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4815..5474)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5389..5408
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     5636..5654
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5658..5701)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    5704..5725
                     /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 gaaagtacta gatgaacaaa cttgacgtca accagtggtt tcctattgct accactgaag
     1201 atctcccgaa gcgccatgtc tttcatgcca cgttgttggg gcaagaaatg gccatctggc
     1261 gcgatgactc tggttcagtt aatgcttggg agaaccgctg cccgcataga ggattgcggt
     1321 tgacactggg tgctaatacc ggtaacgagt tgcggtgtca gtatcatgga tggacttatg
     1381 aaagcgggac tggtggctgc acttttgtcc cagcccatcg cgatgcacca cccccaaatg
     1441 ccgcgcgggt taatactttt cctgtccgcg aaaagcacgg ctttatctgg acgacattag
     1501 gtcagccgcc aggagagccc atttcaatcc tcgatgacgc tcagcttgta aacgctgtaa
     1561 aaacaaatct gcatagcgta gttatagatg ctgatattga cggagttgtc agcgtcctac
     1621 gtcagaatct ttcagcgttc atcgatgtgt ttggtgcggc cagcgctgaa gatctgcatt
     1681 tgaaatccat gctgcaagat cgagggattc tggtaacaag atcaggctct attgctattc
     1741 atttttatat gcagcgctca accattagta aatgcgttgt acatgcgcaa gtacttactc
     1801 cgggacgtcc aggatacgaa cttcaaaaga actactcgta tgccatgaac gttatccgca
     1861 gggcagcaga agctgtagct accgacttga ttagcattac agatatcagc gatcagacta
     1921 tcgaaaagct tgaagtcgtt agagaaaaca tgactaaggc tcctccaacc cactatatct
     1981 gcgaagtggt tacgcgtact caagagacag gtgatattaa ctcatactgg ctgaagccta
     2041 tcggctaccc actaccagca ttcagtccag ggatgcacat cagcatcaca acgccggagg
     2101 gtagcattcg acaatattcc ctcgtgaacg ggcctgacga gcgtgaatcc ttcatcatcg
     2161 gtgtgaagaa agagattcag tcccgtggcg gctccagatc aatgcacgaa gatgtgaagg
     2221 ttggaacgca actaaaagtt acacttccga ggaacggttt tccactcgtc caaaccagaa
     2281 aacacccgat tctcgtagca ggtggcatcg gtatcacccc aattttgtgt atggcacagg
     2341 ctctggatca gcaaggttca tcgtatgaaa tacattattt tgctcgtgca tttgagcatg
     2401 ttccattcca ggatcgactg actgcgttgg gcgatcgttt gaatgtgcat cttggcctcg
     2461 gcccagacga gactagagca aaacttgccg acatcatgga gattcataac gcccaagacg
     2521 tagatgttta cacttgcggc ccgcaaccaa tgatcgaaac tgtatctgct gtcgctcttg
     2581 ctcatggcat cgctgaagag tccatccgat ttgaattttt cagtaaaaag aacgatgttc
     2641 ccgtttctga tgaagaatat gaggttgagc tcaaaaaaac tggtcaaata ttcactgtct
     2701 cgcctggctc tacgttgttg caagcttgtt tggacaacga tgttcgtatc gaagcttctt
     2761 gtgagcaggg tgtatgcggg acttgtataa ctccagtcgt atccggcgat ctcgagcatc
     2821 atgacactta cctttctaag aaagaaaggg aaagcggtaa gtggatcatg ccgtgtgttt
     2881 cgcgctgcaa gtccaaaaaa atcgttctcg atctgtgagg ccgggcatca cccacggtat
     2941 ggacgagctg tataaactag agaaagagga gaaatactag atgatcacgc tttatgacta
     3001 tgaactctcg gggaattgct acaaggtgcg gatgctcctt tccatcctga agctaccgta
     3061 cacgatagag acagtggaat ttttcccgtc cagggaacat aaatcaccgg ccttcctgcg
     3121 catcaatcca ctcggccagt tgccggtatt gcgcgatgat gatttcgtgt tgcgcgatgc
     3181 ccaggccatc ctggtttacc tggcgaacaa atacgacacc agtttgacct ggtatccgac
     3241 acatgcgccc gccatcctcg gccaggcaaa tatgtggctc gcgtttgccg atggtttgac
     3301 cggctcgatt tccgctgcac gtttgcatga tttgtttttt tacgatttca atgcagagca
     3361 atgccgtgcc cgcgcgcacg aactactgcg catactggac gaacacctgt gggcggcgga
     3421 acaaaatgga catcaatggc tatgtccggc cgccgcaccc accatcgcgg atcttgcctg
     3481 cttcccgtat atcgcattgt cggatgaagc cggcgtttca ctactcgact acccggcggt
     3541 acgacgctgg ctggaccgtg tcaaacgcat ccccggattc accgtcatgt cggggatttt
     3601 tccaacctca gcttccttcg atccggcctc acttgcggaa gaaacctgtc aataatacta
     3661 gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc cctttttctt
     3721 taaaaccgaa aagattactt cgcgttatgc aggcttcctc gctcactgac tcgctgcgct
     3781 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3841 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3901 accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct ccgcccccct gacgagcatc
     3961 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4021 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4081 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4141 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4201 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4261 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4321 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4381 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4441 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4501 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4561 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4621 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4681 ctgacagctc gaggcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     4741 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     4801 agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     4861 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     4921 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     4981 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     5041 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     5101 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     5161 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     5221 catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg
     5281 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     5341 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     5401 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     5461 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     5521 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg cccgatcaac
     5581 tcgagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5641 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
     5701 ctggaattcg cggccgcttc tagag
//
