LOCUS       Exported                4110 bp ds-DNA     circular SYN 22-OCT-2019
DEFINITION  Plasmid for regulatable expression of msGFP2 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4110)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Oct 22, 2019 from SnapGene 5.0.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4110
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        198..242
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
                     /note="This forward directional feature has 4 segments:
                        1: 198 .. 212 / #ffffff
                        2: 213 .. 218 / #ffffff / -35
                        3: 219 .. 235 / #ffffff
                        4: 236 .. 242 / #ffffff / -10"
     protein_bind    218..234
                     /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)."
                     /note="color: #31849b"
     protein_bind    250..266
                     /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)."
                     /note="color: #31849b"
     RBS             285..296
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
                     /note="color: #a6acb3"
     CDS             303..1010
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP"
                     /label=msGFP2
                     /note="color: #05fd14"
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     terminator      1044..1138
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
                     /note="color: #ffffff"
     terminator      1910..1996
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     rep_origin      complement(2478..3066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(3237..4097)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3237 .. 4028 / #ccffcc
                        2: 4029 .. 4097 / #ccffcc / signal sequence
                     Cleavage site after base 4028"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(join(4098..4110,1..92))
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
       61 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgct
      121 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
      181 gtcttcacct cgagaaatca taaaaaattt atttgctttg tgagcggata acaattataa
      241 tagattcaat tgtgagcgga taacaatttc acacagaatt cattaaagag gagaaattaa
      301 ccatggatag cactgagagc ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      361 gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg cgagggcgat gccaccaacg
      421 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      481 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
      541 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcacct
      601 tcaaggacga cggcacctac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      661 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
      721 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
      781 gcatcaaggc caacttcaag atccgccaca acgtggagga cggcagcgtg cagctcgccg
      841 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
      901 acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
      961 tgctggagtt cgtgaccgcc gccgggatca ctggaggcag tggcggttct taaagcggcc
     1021 gcaagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga cctcagaact
     1081 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc
     1141 aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     1201 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     1261 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     1321 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     1381 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     1441 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     1501 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     1561 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     1621 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     1681 tcgcccccgt tttcaccatg catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     1741 ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt
     1801 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
     1861 tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg
     1921 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1981 cctgagtagg acaaatccgc cgctctagag ctgcctcgcg cgtttcggtg atgacggtga
     2041 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2101 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     2161 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     2221 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     2281 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2341 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2401 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2461 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2521 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2581 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2641 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2701 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2761 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2821 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2881 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     2941 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3001 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3061 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3121 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3181 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3241 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3301 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3361 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3421 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3481 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3541 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3601 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3661 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3721 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3781 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3841 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3901 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3961 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4021 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4081 aaatgttgaa tactcatact cttccttttt
//
