LOCUS       Exported                7021 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDCAF2, complete sequence.
ACCESSION   KC619529
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7021)
  AUTHORS   Quandt EM, Hammerling MJ, Summers RM, Otoupal PB, Slater B, Alnahhas
            RN, Dasgupta A, Bachman JL, Subramanian MV, Barrick JE.
  TITLE     Decaffeination and measurement of caffeine content by addicted E. 
            coli with a refactored N-demethylation operon
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7021)
  AUTHORS   Quandt EM, Hammerling MJ, Summers RM, Otoupal PB, Slater B, Alnahhas
            RN, Dasgupta A, Bachman JL, Subramanian MV, Barrick JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2013) Chemistry and Biochemistry, The University 
            of Texas at Austin, 2500 Speedway A4800, Austin, TX 78712-1639, USA
REFERENCE   3  (bases 1 to 7021)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7021
                     /organism="Cloning vector pDCAF2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1334236"
     regulatory      22..56
                     /regulatory_class="promoter"
                     /gene="ndmA"
                     /note="BBa_J23100; derived from BBa_K515105"
     regulatory      65..89
                     /regulatory_class="ribosome_binding_site"
                     /note="derived from BBa_K515105"
     RBS             74..82
                     /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)"
     CDS             90..1145
                     /codon_start=1
                     /gene="ndmA"
                     /product="ndmA"
                     /label=ndmA
                     /protein_id="AGL91154.1"
                     /translation="MEQAIINDEREYLRHFWHPVCTVTELEKAHPSSLGPLAVKLLNEQ
                     LVVAKLGDEYVAMRDRCAHRSAKLSLGTVSGNRLQCPYHGWQYDTHGACQLVPACPNSP
                     IPNKAKVDRFDCEERYGLIWIRLDSSFDCTEIPYFSAANDPRLRIVIQEPYWWDATAER
                     RWENFTDFSHFAFIHPGTLFDPNNAEPPIVPMDRFNGQFRFVYDTPEDMAVPNQAPIGS
                     FSYTCSMPFAINLEVSKYSSSSLHVLFNVSCPVDSHTTKNFLIFAREQSDDSDYLHIAF
                     NDLVFAEDKPVIESQWPKDAPADEVSVVADKVSIQYRKWLRELKEAHKEGSQAFRSALL
                     DPVIESDRSYI"
     gene            90..1145
                     /gene="ndmA"
                     /label=ndmA
     regulatory      1153..1164
                     /regulatory_class="ribosome_binding_site"
                     /note="BBa_B0034"
     RBS             1153..1164
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1171..2238
                     /codon_start=1
                     /gene="ndmB"
                     /product="ndmB"
                     /label=ndmB
                     /protein_id="AGL91153.1"
                     /translation="MKEQLKPLLEDKTYLRHFWHPVCTLNEFERANASGHGPMGVTLLG
                     EKLVLARLNSKIIAAADRCAHRSAQLSIGRVCSNAGKDYLECPYHGWRYDEAGACQLIP
                     ACPDKSISPRAKISSFDCEVKYDIVWVRLDNSFDCTQIPYLSDFDNPDMQVIVADSYIW
                     ETVAERRWENFTDFSHFAFVHPGTLYDPFFASHPTVYVNRVDGELQFKLAPPREMKGIP
                     PEAPMGDFTYRCTMPYSVNLEIKLWKDDSRFVLWTTASPVDNKSCRNFMIIVREKDNQP
                     DHMHLAFQKRVLDEDQPVIESQWPLEIQTSEVSVATDKISVQFRKWHKELSLSAVEGRE
                     AFRDSVLTNVIEEEQ"
     gene            1171..2238
                     /gene="ndmB"
                     /label=ndmB
     regulatory      2256..2267
                     /regulatory_class="ribosome_binding_site"
                     /note="BBa_B0034"
     RBS             2256..2267
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2274..3128
                     /codon_start=1
                     /gene="ndmC"
                     /product="ndmC"
                     /label=ndmC
                     /protein_id="AGL91152.1"
                     /translation="MSTDQVIFNDWHPVAALEDVSLDKRYRCRLLGRTVSYVKTSDAVN
                     AHWEESADEIKTIRAKEIYGLLWLSFADKPSEMFDIAEFKEPDRRIVSAGSVRVNVSGL
                     RAIENFLDMAHFPFVHTDILGAEPLTEVEPYNVNYDETVDEIFATECKFPQPKGSATAV
                     EPIDMQYIYRITRPYSAILYKTCPPEPHRWDALGLFIQPVDEDWCIAHTIMCYVDDVNS
                     DQQLRHFQQTIFGQDLMILINQVPKRLPLAASRESPVRADVLATAYRRWLREKGVQYGA
                     LRD"
     gene            2274..3128
                     /gene="ndmC"
                     /label=ndmC
     regulatory      3136..3147
                     /regulatory_class="ribosome_binding_site"
                     /note="BBa_B0034"
     RBS             3136..3147
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3154..4920
                     /codon_start=1
                     /gene="ndmD"
                     /product="ndmD"
                     /label=ndmD
                     /protein_id="AGL91151.1"
                     /translation="MNKLDVNQWFPIATTEDLPKRHVFHATLLGQEMAIWRDDSGSVNA
                     WENRCPHRGLRLTLGANTGNELRCQYHGWTYESGTGGCTFVPAHRDAPPPNAARVNTFP
                     VREKHGFIWTTLGQPPGEPISILDDAQLVNAVKTNLHSVVIDADIDGVVSVLRQNLSAF
                     IDVFGAASAEDLHLKSMLQDRGILVTRSGSIAIHFYMQRSTISKCVVHAQVLTPGRPGY
                     ELQKNYSYAMNVIRRAAEAVATDLISITDISDQTIEKLEVVRENMTKAPPTHYICEVVT
                     RTQETGDINSYWLKPIGYPLPAFSPGMHISITTPEGSIRQYSLVNGPDERESFIIGVKK
                     EIQSRGGSRSMHEDVKVGTQLKVTLPRNGFPLVQTRKHPILVAGGIGITPILCMAQALD
                     QQGSSYEIHYFARAFEHVPFQDRLTALGDRLNVHLGLGPDETRAKLADIMEIHNAQDVD
                     VYTCGPQPMIETVSAVALAHGIAEESIRFEFFSKKNDVPVSDEEYEVELKKTGQIFTVS
                     PGSTLLQACLDNDVRIEASCEQGVCGTCITPVVSGDLEHHDTYLSKKERESGKWIMPCV
                     SRCKSKKIVLDL"
     gene            3154..4920
                     /gene="ndmD"
                     /label=ndmD
     misc_feature    4973..4993
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4994..5051
                     /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."
     rep_origin      complement(5246..5834)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      6017..6111
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6132..6791)
                     /codon_start=1
                     /gene="chlR"
                     /product="chloramphenicol resistance protein"
                     /label=chlR
                     /protein_id="AGL91150.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(6132..6791)
                     /gene="chlR"
                     /label=chlR
     terminator      6975..7018
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7021..21
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""