LOCUS       Exported                7225 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Plasmid vector pCCB9, complete sequence.
ACCESSION   KM219836
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7225)
  AUTHORS   Fernandez-Nino MA, Arevalo-Ferro C, Dreyer I.
  TITLE     A synthetic multi-cellular network of coupled oscillators
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7225)
  AUTHORS   Fernandez-Nino MA, Arevalo-Ferro C, Dreyer I.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JUL-2014) Biologia, Universidad Nacional de Colombia, 
            Avenida Carrera 30 # 45, Bogota, Cundinamarca 111321, Colombia
REFERENCE   3  (bases 1 to 7225)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7225
                     /organism="Plasmid vector pCCB9"
                     /mol_type="other DNA"
                     /db_xref="taxon:1641897"
     rep_origin      180..861
                     /label=ColE1origin
                     /note="ColE1origin"
     rep_origin      complement(198..786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(961..1731)
                     /codon_start=1
                     /product="KanR"
                     /label=KanR
                     /note="kanamycin resistance gene"
                     /protein_id="AKE33862.1"
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG
                     QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
                     SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     terminator      1786..1872
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1916..1932)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1936..1991
                     /regulatory_class="promoter"
                     /note="luxbox"
     CDS             1998..2618
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /note="repressor"
                     /protein_id="AKE33861.1"
                     /translation="MTIMIKKSDFLAIPSEEYKGILSLRYQVFKQRLEWDLVVENNLES
                     DEYDNSNAEYIYACDDTENVSGCWRLLPTTGDYMLKSVFPELLGQQSAPKDPNIVELSR
                     FAVGKNSSKINNSASEITMKLFEAIYKHAVSQGITEYVTVTSTAIERFLKRIKVPCHRI
                     GDKEIHVLGDTKSVVLSMPINEQFKKAVLNSRAANDENYALAA"
     terminator      2633..2719
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2763..2779)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2782..3651
                     /codon_start=1
                     /product="AIIB"
                     /label=AIIB
                     /note="aiiB lactonase"
                     /protein_id="AKE33863.1"
                     /translation="MGNKLFVLDLGEIRVDENFIIANSTFVTPQKPTVSSRLIDIPVSA
                     YLIQCTDATVLYDTGCHPECMGTNGRWPAQSQLNAPYIGASECNLPERLRQLGLSPDDI
                     STVVLSHLHNDHAGCVEYFGKSRLIAHEDEFATAVRYFATGDHSSPYIVKDIEAWLATP
                     RNWDLVGRDERERELAPGVNLLNFGTGHASGMLGLAVRLEKQPGFLLVSDACYTATNYG
                     PPARRAGVLHDTIGYDRTVSHIRQYAESRSLTVLFGHDREQFASLIKSTDGFYESRAAN
                     DENYALAA"
     terminator      3666..3752
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    3733..3815
                     /note="green fluorescent protein; GFP"
     primer_bind     complement(3796..3812)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3867..3953
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3997..4013)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4020..4724
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELY"
     terminator      4784..4870
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4914..4930)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4937..5032
                     /regulatory_class="promoter"
                     /note="Pllac01"
     protein_bind    4940..4956
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    4963..4979
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5021..5032
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5039..5827
                     /codon_start=1
                     /product="LuxR"
                     /label=LuxR
                     /note="LuxR receptor"
                     /protein_id="AKE33864.1"
                     /translation="MKNINADDTYRIINKIKACRSNNDINQCLSDMTKMVHCEYYLLAI
                     IYPHSMVKSDISILDNYPKKWRQYYDDANLIKYDPIVDYSNSNHSPINWNIFENNAVNK
                     KSPNVIKEAKSSGLITGFSFPIHTANNGFGMLSFAHSEKDNYIDSLFLHACMNIPLIVP
                     SLVDNYRKINIANNKSNNDLTKREKECLAWACEGKSSWDISKILGCSKRTVTFHLTNAQ
                     MKLNTTNRCQSISKAILTGAIDCPYFKNSAANDENYALAA"
     terminator      5842..5928
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5972..5988)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5994..6608
                     /codon_start=1
                     /product="LuxI"
                     /label=LuxI
                     /note="LuxI AHL synthase"
                     /protein_id="AKE33865.1"
                     /translation="MTIMIKKSDFLAIPSEEYKGILSLRYQVFKQRLEWDLVVENNLES
                     DEYDNSNAEYIYACDDTENVSGCWRLLPTTGDYMLKSVFPELLGQQSAPKDPNIVELSR
                     FAVGKNSSKINNSASEITMKLFEAIYKHAVSQGITEYVTVTSTAIERFLKRIKVPCHRI
                     GDKEIHVLGDTKSVVLSMPINEQFKKAVLNAANDENYALAA"
     terminator      6623..6709
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6753..6769)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6971..7075)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      7165..7208
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"