pLEVI(408)-ColE-iLight-msfGFP

LOCUS       pLEVI(408)-ColE-iLight-msfGFP                7703 bp ds-DNA     circular SYN 15-JUN-2021
DEFINITION  Expresses LexA_DBD-iLight-msfGFP under J23116 promoter and mCherry 
            under ColE promoter with a LexA408 operator in bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    pLEVI(408)-ColE-iLight-msfGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7703)
  TITLE     Single-component near-infrared optogenetic systems for gene 
            transcription regulation
REFERENCE   2  (bases 1 to 7703)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7703
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1958..2677
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2003..2024)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2611..2632
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2887..2933
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2956..2974)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2970..3017
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3188..3979)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3980..4071)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4119..4857)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(5067..6149)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6111..6130
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6150..6227)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      6453..6499
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             6675..7385
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6820..6838)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7088..7107
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      7592..7638
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cagctagctc agtcctaggg actatgctag caaacactat aacttcctag gtattattat
       61 gaaagcgtta acggccaggc aacaagaggt gtttgatctc atccgtgatc acatcagcca
      121 gacaggtatg ccgccgacgc gtgcggaaat cgcgcagcgt ttggggttcc gttccgcgtc
      181 ctcggctgaa gaacatctga aggcgctggc acgcaaaggc gttattgaaa ttgtttccgg
      241 cgcatcacgc gggattcgtc tgttgcagga agaggaagaa gggttgccgc tggtaggtcg
      301 tgtggctgcc ggtgaaccgc ggagtgctgg tggggcagca gatctgggta gtgatgatat
      361 cagcaaactg attgcagcat gtgatcaaga accgattcat attccgaatg caattcagcc
      421 gtttggtgca atgctgattg ttgaaaaaga tacccagcag attgtttatg caagcgcaaa
      481 tagcgcagaa tatttcagcg ttgcagataa caccattcat gaactgagcg attttaaaca
      541 ggccaacatt aatagcctgc tgccggaaca actgattagc ggtctgacaa gcgcaattag
      601 tgaaaatgaa ccgatttggg ttgaaaccga tcgtctgagc tttctgggtt ggcgtcatga
      661 aaactattac atcattgaag tggaacgcta tcatgtgcag accagcaatt ggtttgaaat
      721 tcagtttcag cgtgcctttc agaaactgcg taattgcaaa acccataacg atctgattaa
      781 taccctgacc cgtctgattc aagaaatcag cggttatgat cgcgtgatga tctatcaatt
      841 tgatccggaa tggaatggtc gtgttattgc agaaagcgtt cgtcagctgt ttaccagcat
      901 gctgaatcat cattttccgg caagcgatat tccggcacag gcacgtgcaa tgtatagcat
      961 taatccgatt cgtattatcc cggatgttaa tgcagaaccg cagccgctgc acatgattca
     1021 taaaccgcaa aataccgaag cagttaatct gagctgcggt gttctgcgtg cagttagccc
     1081 tctgcacatg cagtatctgc gtaattttgg tgttagcgca agcaccagca ttggcatttt
     1141 taacgaagat aaactgtggg gtatcgttgc atgccatcat accaaaccgc gtgcaattgg
     1201 tcgtcgtatt cgtcatctgc tggttcgtac cgttgaattt gcagcagaac gtctgtggct
     1261 gattcatagc cgtaatgttg aacgttatat ggttaccgtt caggcagcac gtgaacagct
     1321 gagcaccacc gcagatgata aacattcaag ccatgaaatc gtgattgaac atgcagcaga
     1381 ttggtgtaaa ctgtttcgtt gtgatggtgt tggttatctg cgtggagaag aactgaccac
     1441 ctatggtgaa acaccggatc agaccaccat taacaaactg gttgaatggc tggaagagaa
     1501 cggtaaaaaa agcctgtttt ggcatagcca catgctgaaa gaagatgcac cgggtctgct
     1561 gccggatggt agccgttttg caggtctgct ggcaattccg ctgaaaagtg atgcagacct
     1621 gtttagctat ctgctgctgt ttcgtgtggc acagaatgaa gttcgtacct gggcgggtaa
     1681 accggaaaaa ctgagcgttg aaaccagcac tggcaccatg gtgggtccgc gtaaaagttt
     1741 tgaagcatgg caggatgaag ttagcggtaa aagccagccg tggcgtaccg cacagctgta
     1801 tgcagcacgt gatattgcac gtgatctgct gattgtggca gatagcatgc agctgaatct
     1861 gctgaatgat cagctggcag atgcaaatga aaatctggaa aaactggcca gctttgatga
     1921 tctgaccgtc gactccggtg gtggttctgg tggtggaatg gtgagcaagg gcgaggagct
     1981 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2041 cagcgtgcgc ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat
     2101 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2161 cgtgcagtgc ttcagccgct accccgacca catgaagcgc cacgacttct tcaagtccgc
     2221 catgcccgaa ggctacgtcc aggagcgcac catcagcttc aaggacgacg gcacctacaa
     2281 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2341 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag
     2401 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     2461 ccgccacaac gtggaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2521 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     2581 gagcaaagat cccaacgaga aacgcgatca catggtcctg ctggagttcg tgaccgccgc
     2641 cgggatcact ctcggcatgg acgagctgta caagtgagaa ttccccctgt tttggcggat
     2701 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     2761 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     2821 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     2881 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     2941 ttgctcgagc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     3001 tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc
     3061 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
     3121 ctgaaccgac gaccgggtca tcgtggccgg atcttgcggc ccctcggctt gaacgaattg
     3181 ttagacatta tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt
     3241 ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag
     3301 cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga
     3361 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca
     3421 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat
     3481 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac
     3541 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga
     3601 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca
     3661 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt
     3721 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga
     3781 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat
     3841 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg
     3901 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg
     3961 cgatcaccgc ttccctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4021 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa atagctagct
     4081 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
     4141 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
     4201 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
     4261 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
     4321 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
     4381 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
     4441 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
     4501 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
     4561 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
     4621 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
     4681 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
     4741 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
     4801 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4861 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     4921 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
     4981 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     5041 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5101 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     5161 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     5221 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     5281 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     5341 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     5401 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     5461 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     5521 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     5581 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     5641 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     5701 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     5761 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     5821 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     5881 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     5941 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     6001 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     6061 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     6121 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     6181 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     6241 ctcggtaccc ccctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc
     6301 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
     6361 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
     6421 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     6481 actgggcctt tcgttttatc tgttgtttgc gctagcggat cctgtttttt tgatcgtttt
     6541 cacaaaaatg gaagtacaca gtcttgacag ggaaaatgca gcggcgtagc ttttatgccg
     6601 tatataaaaa cggcgtttat atgtacggta tttattttta acttattgtt ttaaaagtca
     6661 aagaggattt tataatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     6721 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     6781 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     6841 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     6901 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     6961 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     7021 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     7081 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     7141 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     7201 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     7261 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     7321 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     7381 agtaaagatc tctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     7441 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     7501 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     7561 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     7621 ctgggccttt cgttttatct gttgtttgcg ctagcggatc cgcctatgca gcgacaaata
     7681 ttgatagcct gaatcagtat tga
//