LOCUS       Exported                5080 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFNK-512, complete sequence.
ACCESSION   MH300675
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5080)
  AUTHORS   Karig D, Martini KM, Lu T, DeLateur NA, Goldenfeld N, Weiss R.
  TITLE     Stochastic Turing patterns in a synthetic bacterial population
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. (2018) In press
  PUBMED    29891706
REFERENCE   2  (bases 1 to 5080)
  AUTHORS   Karig DK, Lu T.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2018) Research and Development Department, Johns 
            Hopkins University Applied Physics Laboratory, 11100 Johns Hopkins 
            Rd, Laurel, MD 20723, USA
REFERENCE   3  (bases 1 to 5080)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5080
                     /organism="Cloning vector pFNK-512"
                     /mol_type="other DNA"
                     /db_xref="taxon:2231587"
     CDS             complement(134..853)
                     /codon_start=1
                     /product="LasR"
                     /label=LasR
                     /protein_id="AWV91933.1"
                     /translation="MALVDGFLELERSSGKLEWSAILQKMASDLGFSKILFGLLPKDSQ
                     DYENAFIVGNYPAAWREHYDRAGYARVDPTVSHCTQSVLPIFWKPSIYQTRKQHEFFEE
                     ASAAGLVYGLTMPLHGARGELGALSLSVEAENRAEANRFMESVLPTLWMLKDYALQSGA
                     GLAFEHPVSKPAVLTSREKEVLQWCAIGKTSWEISVICNCSEANVNFHMGNIRRKFGVT
                     SRRVAAIMAVNLGLITL"
     RBS             860..871
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      complement(863..871)
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS II"
     regulatory      complement(892..964)
                     /regulatory_class="promoter"
                     /note="prokaryotic; qscrsaL-OR1-mut4"
     misc_feature    complement(892..908)
                     /label=cI OR1 mut4
                     /note="cI OR1 mut4"
     misc_feature    complement(941..960)
                     /label=las box - qscrsaL
                     /note="las box - qscrsaL"
     regulatory      1019..1091
                     /regulatory_class="promoter"
                     /note="prokaryotic; qscrsaL-OR1m4"
     misc_feature    1023..1042
                     /label=las box - qscrsaL
                     /note="las box - qscrsaL"
     misc_feature    1075..1091
                     /label=cI OR1 mut4
                     /note="cI OR1 mut4"
     RBS             1098..1109
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      1098..1106
                     /regulatory_class="ribosome_binding_site"
                     /note="RBSII"
     CDS             1116..1760
                     /codon_start=1
                     /product="LasI(LVA)"
                     /label=LasI
                     /protein_id="AWV91934.1"
                     /translation="MIVQIGRREEFDKKPLGEMHKLRAQVFKERKGWDVSVIDEMEIDG
                     YDALSPYYMLIQEDTPEAQVFGCWRILDTTGPYMLKNTFPELLHGKEAPCSPHIWELSR
                     FAINSGQKGSLGFSDCTLEAMRALARYSLQNDIQTLVTVTTVGVEKMMIRAGLDVSRFG
                     PHLKIGIERAVALRIELNAKTQIALYGGVLVEQRLAVSRPAANDENYALVA"
     misc_feature    1719..1757
                     /label=Region: LVA
                     /note="Region: LVA"
     RBS             1824..1835
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      1824..1832
                     /regulatory_class="ribosome_binding_site"
                     /note="RBSII"
     CDS             1842..2447
                     /codon_start=1
                     /product="RhlI"
                     /label=RhlI
                     /protein_id="AWV91935.1"
                     /translation="MIELLSESLEGLSAAMIAELGRYRHQVFIEKLGWDVVSTSRVRDQ
                     EFDQFDHPQTRYIVAMSRQGICGCARLLPTTDAYLLKDVFAYLCSETPPSDPSVWELSR
                     YAASAADDPQLAMKIFWSSLQCAWYLGASSVVAVTTTAMERYFVRNGVILQRLGPPQKV
                     KGETLVAISFPAYQERGLEMLLRYHPEWLQGVPLSMAV"
     RBS             2496..2507
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      2496..2504
                     /regulatory_class="ribosome_binding_site"
                     /note="RBSII"
     CDS             2514..3191
                     /codon_start=1
                     /product="DsRed-Exp"
                     /label=DsRed-Exp
                     /protein_id="AWV91936.1"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     regulatory      3270..3374
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      3274..3360
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      3503..4141
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(3530..4118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      complement(4200..4305)
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      4206..4300
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4321..4980)
                     /codon_start=1
                     /product="Cm(r)"
                     /label=Cm
                     /protein_id="AWV91937.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(4981..3)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"