LOCUS       sequence-148247-        2967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RFP constitutively expressed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2967)
  AUTHORS   Abudayyeh OO, Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox
            DB, Shmakov S, Makarova KS, Semenova E, Minakhin L, Severinov K, 
            Regev A, Lander ES, Koonin EV, Zhang F
  TITLE     C2c2 is a single-component programmable RNA-guided RNA-targeting 
            CRISPR effector.
  JOURNAL   Science. 2016 Jun 2. pii: aaf5573.
  PUBMED    27256883
REFERENCE   2  (bases 1 to 2967)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2016 Jun 2. pii: aaf5573."
COMMENT     Sequence Label: pC007 - pRFP constitutive expression
FEATURES             Location/Qualifiers
     source          1..2967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     137..155
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(159..202)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     protein_bind    220..238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    245..263
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             281..292
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             299..976
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      994..1065
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1081..1108
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1141..1158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1312..1900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1392..1411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2071..2862,2863..2931))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2694..2713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(2932..2967,1..69))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
       61 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
      121 gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt
      181 agctttcgct aaggatgatt tctggaattc atgagatctt ccctatcagt gatagagatt
      241 gacatcccta tcagtgatag agatactgag cacggatctg aaagaggaga aaggatctat
      301 ggcgagtagc gaagacgtta tcaaagagtt catgcgtttc aaagttcgta tggaaggttc
      361 cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac
      421 ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt gggacatcct
      481 gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg acatcccgga
      541 ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga acttcgaaga
      601 cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt tcatctacaa
      661 agttaaactg cgtggtacca acttcccgtc cgacggtccg gttatgcaga aaaaaaccat
      721 gggttgggaa gcttccaccg aacgtatgta cccggaagac ggtgctctga aaggtgaaat
      781 caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta aaaccaccta
      841 catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca aactggacat
      901 cacctcccac aacgaagact acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca
      961 ctccaccggt gcttaataag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa
     1021 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     1081 ggctcacctt cgggtgggcc tttctgcgtt tataggatcc taactcgagc tgcaggcttc
     1141 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1201 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1261 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1321 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1381 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1441 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1501 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1561 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1621 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1681 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1741 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1801 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1861 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1921 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1981 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2041 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2101 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2161 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2221 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2281 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2341 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt
     2401 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2461 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2521 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2581 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2641 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     2701 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2761 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2821 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2881 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2941 ttttcaatat tattgaagca tttatca
//
