LOCUS       sequence-489194-        4431 bp    DNA     circular SYN 14-OCT-2025
DEFINITION  Reporter J1-pspAp-mRFP with 1bp deletion phase-shift.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4431)
  AUTHORS   Kiattisewee C, Karanjia AV, Cardiff RAL, Olander KE, Leejareon P, 
            Alvi SS, Carothers JM, Zalatan JG
  TITLE     Systematic Mapping of Bacterial CRISPRa Systems for Synergistic Gene
            Activation Reveals Antagonistic Effects.
  JOURNAL   ACS Synth Biol. 2025 Jul 22. doi: 10.1021/acssynbio.5c00358.
  PUBMED    40693287
REFERENCE   2  (bases 1 to 4431)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2025 Jul 22. doi: 10.1021/acssynbio.5c00358."
COMMENT     Sequence Label: pCK662.-1
FEATURES             Location/Qualifiers
     source          1..4431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     619..638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(857..961)
                     /gene="bla"
                     /label=AmpR promoter
     RBS             1218..1229
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1236..1913
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1946..2017
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2033..2060
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2591..3541)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(4303..4397)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(4427..4431,1..787,788..856))
                     /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"
ORIGIN
        1 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
       61 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      121 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
      181 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      241 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
      301 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      361 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      421 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      481 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
      541 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
      601 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
      661 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
      721 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
      781 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
      841 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
      901 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
      961 gcacatttcc ccgaaaagtg ccacctgtgg caattccgac gtcgcctacg gtatccaccg
     1021 gagacctatg gcagcctccg gccgccatag gacacctttg gttgccaagg gtgacctatg
     1081 gtgaccatgg gccaccacgg gcgacctcag gtatcctgcg gtgtcctgcg gttaccaaag
     1141 gcgtcctttg ggttccaccg gatacctcgg acataaaaaa ttggcacgca aattgtatta
     1201 acagttcaga attcattaaa gaggagaaag gtaccatggc gagtagcgaa gacgttatca
     1261 aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac gagttcgaaa
     1321 tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta
     1381 ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc cccgcagttc cagtacggtt
     1441 ccaaagctta cgttaaacac ccggctgaca tcccggacta cctgaaactg tccttcccgg
     1501 aaggtttcaa atgggaacgt gttatgaact tcgaagacgg tggtgttgtt accgttaccc
     1561 aggactcctc cctgcaagac ggtgagttca tctacaaagt taaactgcgt ggtaccaact
     1621 tcccgtccga cggtccggtt atgcagaaaa aaaccatggg ttgggaagct tccaccgaac
     1681 gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag
     1741 acggtggtca ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc
     1801 tgccgggtgc ttacaaaacc gacatcaaac tggacatcac ctcccacaac gaagactaca
     1861 ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc caccggtgct taaggatcca
     1921 aactcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     1981 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     2041 ttcgggtggg cctttctgcg tttataccta gggtacgggt tttgctgccc gcaaacgggc
     2101 tgttctggtg ttgctagttt gttatcagaa tcgcagatcc ggcttcagcc ggtttgccgg
     2161 ctgaaagcgc tatttcttcc agaattgcca tgattttttc cccacgggag tcgtcactgg
     2221 ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc gcctgtttca ggctgtctat
     2281 gtgtgactgt tgagctgtaa caagttgtct caggtgttca atttcatgtt ctagttgctt
     2341 tgttttactg gtttcacctg ttctattagg tgttacatgc tgttcatctg ttacattgtc
     2401 gatctgttca tggtgaacag ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc
     2461 tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg atatgtaacg
     2521 gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc
     2581 cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt
     2641 tttgcgtgag ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa
     2701 attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc
     2761 ttagtccgtt atgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca
     2821 tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt
     2881 ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt
     2941 aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat
     3001 ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg
     3061 ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt
     3121 atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga
     3181 aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg
     3241 catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag
     3301 ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga
     3361 tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag
     3421 ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct
     3481 ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca
     3541 tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat
     3601 gataattact agtccttttc ccgggtgatc tgggtatctg taaattctgc tagacctttg
     3661 ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt
     3721 tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa
     3781 aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca
     3841 aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct
     3901 taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc
     3961 aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca
     4021 gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc
     4081 ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt
     4141 ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca
     4201 cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt
     4261 atcatcaaca ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa
     4321 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     4381 atctatcaac aggagtccaa gcgagctcgt aaacttggtc tgacagttac c
//
