LOCUS       sequence-409055-        3504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J23107.MCP-GAI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3504)
  AUTHORS   Alba Burbano D, Cardiff RAL, Tickman BI, Kiattisewee C, Maranas CJ, 
            Zalatan JG, Carothers JM
  TITLE     Engineering activatable promoters for scalable and multi-input 
            CRISPRa/i circuits.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Jul 25;120(30):e2220358120. doi: 
            10.1073/pnas.2220358120. Epub 2023 Jul 18.
  PUBMED    37463216
REFERENCE   2  (bases 1 to 3504)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-07-25"; volume: "120(30):e2220358120. 
            doi"; pages: " 10.1073/pnas.2220358120. Epub 2023 Jul 18"
COMMENT     Sequence Label: pRC042
FEATURES             Location/Qualifiers
     source          1..3504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(20..608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(100..119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(779..1570,1571..1639))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1402..1421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1640..1744)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1812..1830
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2182..2216
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     RBS             2226..2237
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2628..2645
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3353..3370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aggccgcgtt gctggcgttt ttccataggc tccgcacccc tgacgagcat cacaaaaatc
       61 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      121 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      181 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      241 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      301 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      361 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      421 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      481 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      541 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      601 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      661 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      721 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      781 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
      841 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      901 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
      961 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1021 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1081 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1141 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1201 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1261 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1321 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1381 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1441 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1501 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1561 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1621 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1681 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1741 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1801 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc tggcgaatcc
     1861 tctgaccagc cagaaaacga cctttctgtg gtgaaaccgg atgctgcaat tcagagcgcc
     1921 agcaagtggg ggacagcaga agacctgacc gccgcagagt ggatgtttga catggtgaag
     1981 actatcgcac catcagccag aaaaccgaat tttgctgggt gggctaacga tatccgcctg
     2041 atgcgtgaac gtgacggacg taaccaccgc gacatgtgtg tgctgttccg ctgggcatgc
     2101 caggacaact tctggtccgg taacgtgctg agcccggcca agcttagccc gaaaagtgcc
     2161 acctgtggca attccgacgt ctttacggct agctcagccc taggtattat gctagcgaat
     2221 tcattaaaga ggagaaaggt accatggggc ccgcttctaa ctttactcag ttcgttctcg
     2281 tcgacaatgg cggaactggc gacgtgactg tcgccccaag caacttcgct aacgggatcg
     2341 ctgaatggat cagctctaac tcgcgttcac aggcttacaa agtaacctgt agcgttcgtc
     2401 agagctctgc gcagaatcgc aaatacacca tcaaagtcga ggtgcctaaa ggcgcctggc
     2461 gttcgtactt aaatatggaa ctaaccattc caattttcgc cacgaattcc gactgcgagc
     2521 ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc tcagcaatcg
     2581 cagcaaactc cggcatctac ggtggcggag gtagcatgaa gcgcgatcat catcaccacc
     2641 accaccagga taaaaagacg atgatgatga atgaggaaga tgatggaaac gggatggacg
     2701 aattgctggc agtgctggga tataaggtgc gttcgtccga aatggcagat gttgctcaga
     2761 aattggagca gttagaagta atgatgagta acgttcaaga agatgatctt tcacagttag
     2821 cgaccgaaac tgtccactac aaccctgctg agctttacac ttggttggac tccatgctta
     2881 ccgatcttaa ctgacgcaaa aaaccccgct tcggcggggt tttttcgcgg atccttactc
     2941 cgtcgacaag cttgcggccg cactcgagga attccgactc aattagttca gtcagtttca
     3001 ggatattagt catctctaca ttgattatga gtattcagaa attccttaaa tattctgaca
     3061 aatgctcttt ccctaaactc cccccataaa aaaacccgcc gaagcgggtt tttacgttat
     3121 ttgcggatta acgattactc gttatcagaa ccgcccagac ctgcgttcag cagttctgcc
     3181 aggctggcag atgcgtcttc cggaattgat ccgtcgaccg atgcccttga gagccttcaa
     3241 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     3301 cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca
     3361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3481 agcaaaaggc caggaaccgt aaaa
//
