LOCUS       40924_18266        4223 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pET-SP6p-lacO-GFP-LVA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4223)
  AUTHORS   Schaerli Y, Munteanu A, Gili M, Cotterell J, Sharpe J, Isalan M.
  TITLE     A unified design space of synthetic stripe-forming networks
  JOURNAL   Nat Commun 5, 4905 (2014)
  PUBMED    25247316
REFERENCE   2  (bases 1 to 4223)
  AUTHORS   Schaerli Y, Munteanu A, Gili M, Cotterell J, Sharpe J, Isalan M.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JUL-2014) Centre for Genomic Regulation (CRG), 
            EMBL/CRG Systems Biology Research Unit, Dr. Aiguader 88, Barcelona, 
            Barcelona 08003, Spain
REFERENCE   3  (bases 1 to 4223)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4223)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            5, 4905 (2014)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JUL-2014) Centre for Genomic Regulation (CRG), EMBL/CRG Systems 
            Biology Research Unit, Dr. Aiguader 88, Barcelona, Barcelona 08003, 
            Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             495..683
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    788..930
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1116..1704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1878..2735)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2736..2840)
                     /label=AmpR promoter
     primer_bind     2880..2896
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2909..2927
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    2929..2953
                     /label=lac operator
                     /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             3061..3083
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3091..3804
                     /codon_start=1
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3954..4001
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4010..4096
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4188..4215
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 agatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga
       61 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
      121 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
      181 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta
      241 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
      301 taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg
      361 ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta
      421 tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt
      481 acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
      541 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
      601 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt
      661 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
      721 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
      781 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
      841 tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac
      901 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
      961 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1021 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1081 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1141 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1201 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1261 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1321 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1381 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1441 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1501 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     1561 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1621 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1681 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1741 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     1801 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1861 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     1921 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     1981 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2041 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2101 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2161 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     2221 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2281 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2341 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2401 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2461 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     2521 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     2581 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     2641 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     2701 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     2761 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2821 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cctcgagttg
     2881 taaaacgacg gccagtcaat tggaattcat ttaggtgaca ctatagaagg aattgtgagc
     2941 ggataacaat tccgagctcg ttgagttacc tgcagcgatt gtactagttg cggccgcctg
     3001 cgcaccacct gcaggacctc agcaatcgct tggaccagct ttcttaaggg taccaataat
     3061 tttgtttaac tttaagaagg agatatacat atgcgtaaag gcgaagaact gtttaccggt
     3121 gtggttccga ttctggtgga actggacggc gatgttaatg gtcataaatt cagtgttcgc
     3181 ggcgaaggtg aaggcgatgc gacgaacggc aaactgaccc tgaaatttat ctgcaccacg
     3241 ggtaaactgc cggtcccgtg gccgacgctg gtgaccacgc tgacctatgg cgttcaatgt
     3301 tttgcgcgtt acccggatca catgaaacag cacgactttt tcaaatcggc catgccggaa
     3361 ggctatgtgc aggaacgtac gattagcttt aaagacgatg gtacgtataa aacccgcgcg
     3421 gaagtgaaat tcgaaggcga taccctggtt aaccgtatcg aactgaaagg tatcgatttc
     3481 aaagaagacg gcaatattct gggtcataaa ctggaatata acttcaattc ccacaacgtg
     3541 tacatcaccg cggataaaca gaaaaacggc attaaagcca atttcaaaat ccgccataat
     3601 gtggaagatg gtagcgttca gctggccgac cactatcagc aaaacacgcc gattggtgat
     3661 ggcccggtcc tgctgccgga caatcactac ctgagtaccc agtccgtgct gtcaaaagat
     3721 ccgaacgaaa aacgtgacca catggtcctg ctggaatttg tgacggctgc gggtatcacc
     3781 cacggcatgg acgaactgta taaaaggcct gctgctaacg atgaaaacta cgcactggtg
     3841 gcttaataaa ggccttaata aggatccggc gccgctcatt cgctaatcgc cacgacgcgt
     3901 agtcgacccg ccaattccca tggggcgcgc ctgctgccac cgctgagcaa taactagcat
     3961 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gccaggcatc aaataaaacg
     4021 aaaggctcag tcggaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     4081 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg
     4141 gtggcgggca ggacgcccgc cataaactgc caggcatcaa actaagcaga aggccatcct
     4201 gacggatggc ctttttgcgt ttc
//
