LOCUS       sequence-486115-        4553 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Reporter J1-J3(122) -BBa_J23117-mRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4553)
  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 4553)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 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: pAK011
FEATURES             Location/Qualifiers
     source          1..4553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(30..124)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(154..945,946..1014))
                     /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     777..796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1015..1119)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1454..1488
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             1498..1509
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1516..2193
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2226..2297
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2313..2340
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2871..3821)
                     /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 ttacccgtct tactgtccct agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
       61 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
      121 agtccaagcg agctcgtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
      181 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
      241 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
      301 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      361 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
      421 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
      481 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
      541 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
      601 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
      661 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
      721 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
      781 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
      841 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
      901 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
      961 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1021 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1081 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1141 acctgtggca attccgacgt cgcctacggt atccaccgga gacctatggc agcctccggc
     1201 cgccatagga cacctttggt tgccaagggt gacctatggt gaccatgggc caccacgggc
     1261 gacctcaggt atcctgcggt gtcctgcggt taccaaaggc gtcctttggg ttccaccgga
     1321 tacctccgga cgtcataatc atcgctacga gctgtgaaag atgcataaag ctcgtacgac
     1381 gcgttcgctc gtctcctcac ttctcctacg gagcgttctg gacacaacgt cgtcttgaag
     1441 ttgcgattat agattgacag ctagctcagt cctagggatt gtgctagcga attcattaaa
     1501 gaggagaaag gtaccatggc gagtagcgaa gacgttatca aagagttcat gcgtttcaaa
     1561 gttcgtatgg aaggttccgt taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt
     1621 cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg
     1681 ttcgcttggg acatcctgtc cccgcagttc cagtacggtt ccaaagctta cgttaaacac
     1741 ccggctgaca tcccggacta cctgaaactg tccttcccgg aaggtttcaa atgggaacgt
     1801 gttatgaact tcgaagacgg tggtgttgtt accgttaccc aggactcctc cctgcaagac
     1861 ggtgagttca tctacaaagt taaactgcgt ggtaccaact tcccgtccga cggtccggtt
     1921 atgcagaaaa aaaccatggg ttgggaagct tccaccgaac gtatgtaccc ggaagacggt
     1981 gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca ctacgacgct
     2041 gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc
     2101 gacatcaaac tggacatcac ctcccacaac gaagactaca ccatcgttga acagtacgaa
     2161 cgtgctgaag gtcgtcactc caccggtgct taaggatcca aactcgagta aggatctcca
     2221 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2281 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     2341 tttataccta gggtacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt
     2401 gttatcagaa tcgcagatcc ggcttcagcc ggtttgccgg ctgaaagcgc tatttcttcc
     2461 agaattgcca tgattttttc cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct
     2521 ttgattcgat aagcagcatc gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa
     2581 caagttgtct caggtgttca atttcatgtt ctagttgctt tgttttactg gtttcacctg
     2641 ttctattagg tgttacatgc tgttcatctg ttacattgtc gatctgttca tggtgaacag
     2701 ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt
     2761 catctgtgca tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt
     2821 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
     2881 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     2941 gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     3001 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag
     3061 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     3121 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     3181 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     3241 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     3301 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     3361 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     3421 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     3481 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     3541 atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     3601 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     3661 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     3721 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     3781 atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg
     3841 tgattttaat cactatacca attgagatgg gctagtcaat gataattact agtccttttc
     3901 ccgggtgatc tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct
     3961 gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg
     4021 gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct
     4081 gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg
     4141 tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc
     4201 tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct
     4261 ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca
     4321 ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt
     4381 cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc
     4441 tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca
     4501 ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca ggc
//
