LOCUS       sequence-213780-        4431 bp    DNA     circular SYN 14-APR-2025
DEFINITION  J1-mRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4431)
  AUTHORS   Dong C, Fontana J, Patel A, Carothers JM, Zalatan JG
  TITLE     Synthetic CRISPR-Cas gene activators for transcriptional 
            reprogramming in bacteria.
  JOURNAL   Nat Commun. 2018 Jun 27;9(1):2489. doi: 10.1038/s41467-018-04901-6.
  PUBMED    29950558
REFERENCE   2  (bases 1 to 4431)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-06-27"; volume: "9(1):2489. doi"; pages: " 
            10.1038/s41467-018-04901-6"
COMMENT     Sequence Label: pJF076Sa
FEATURES             Location/Qualifiers
     source          1..4431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(112..206)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(236..1027,1028..1096))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     859..878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1097..1201)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1414..1448
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             1458..1469
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1476..2153
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2186..2257
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2273..2300
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2831..3781)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa
       61 ggcagcggta tcatcaacag gcttacccgt cttactgtcc ctagtgcttg gattctcacc
      121 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
      181 cattactgga tctatcaaca ggagtccaag cgagctcgta aacttggtct gacagttacc
      241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
      421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
      541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
      781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
      841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
      901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
      961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1141 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1201 gcacatttcc ccgaaaagtg ccacctgtgg caattccgac gtcgcctacg gtatccaccg
     1261 gagacctatg gcagcctccg gccgccatag gacacctttg gttgccaagg gtgacctatg
     1321 gtgaccatgg gccaccacgg gcgacctcag gtatcctgcg gtgtcctgcg gttaccaaag
     1381 gcgtcctttg ggttccaccg gatacctccg gacttgacag ctagctcagt cctagggatt
     1441 gtgctagcga attcattaaa gaggagaaag gtaccatggc gagtagcgaa gacgttatca
     1501 aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac gagttcgaaa
     1561 tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta
     1621 ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc cccgcagttc cagtacggtt
     1681 ccaaagctta cgttaaacac ccggctgaca tcccggacta cctgaaactg tccttcccgg
     1741 aaggtttcaa atgggaacgt gttatgaact tcgaagacgg tggtgttgtt accgttaccc
     1801 aggactcctc cctgcaagac ggtgagttca tctacaaagt taaactgcgt ggtaccaact
     1861 tcccgtccga cggtccggtt atgcagaaaa aaaccatggg ttgggaagct tccaccgaac
     1921 gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag
     1981 acggtggtca ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc
     2041 tgccgggtgc ttacaaaacc gacatcaaac tggacatcac ctcccacaac gaagactaca
     2101 ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc caccggtgct taaggatcca
     2161 aactcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2221 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     2281 ttcgggtggg cctttctgcg tttataccta gggtacgggt tttgctgccc gcaaacgggc
     2341 tgttctggtg ttgctagttt gttatcagaa tcgcagatcc ggcttcagcc ggtttgccgg
     2401 ctgaaagcgc tatttcttcc agaattgcca tgattttttc cccacgggag gcgtcactgg
     2461 ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc gcctgtttca ggctgtctat
     2521 gtgtgactgt tgagctgtaa caagttgtct caggtgttca atttcatgtt ctagttgctt
     2581 tgttttactg gtttcacctg ttctattagg tgttacatgc tgttcatctg ttacattgtc
     2641 gatctgttca tggtgaacag ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc
     2701 tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg atatgtaacg
     2761 gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc
     2821 cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt
     2881 tttgcgtgag ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa
     2941 attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc
     3001 ttagtccgtt atgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca
     3061 tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt
     3121 ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt
     3181 aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat
     3241 ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg
     3301 ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt
     3361 atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga
     3421 aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg
     3481 catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag
     3541 ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga
     3601 tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag
     3661 ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct
     3721 ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca
     3781 tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat
     3841 gataattact agtccttttc ccgggtgatc tgggtatctg taaattctgc tagacctttg
     3901 ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt
     3961 tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa
     4021 aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca
     4081 aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct
     4141 taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc
     4201 aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca
     4261 gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc
     4321 ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt
     4381 ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc a
//
