LOCUS       sequence-102224-        5130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circuit: OR gate, the sgRNA used for the NOT gate was 
            substituted with one designed to target the malT gene in the E. coli
            genome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5130)
  AUTHORS   Nielsen AA, Voigt CA
  TITLE     Multi-input CRISPR/Cas genetic circuits that interface host 
            regulatory networks.
  JOURNAL   Mol Syst Biol. 2014 Nov 24;10:763. doi: 10.15252/msb.20145735.
  PUBMED    25422271
REFERENCE   2  (bases 1 to 5130)
  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: "Mol Syst 
            Biol."; date: "2014-11-24"; volume: "10:763. doi"; pages: " 
            10.15252/msb.20145735"
COMMENT     Sequence Label: pAN-OR-MalT-3NT
FEATURES             Location/Qualifiers
     source          1..5130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    91..112
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(113..991)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1018..1302
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1225..1244
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        1323..1398
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1419..1448
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1534..1551)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1704..1750
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1803..1820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1974..2562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2054..2073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2733..3524,3525..3593))
                     /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     3356..3375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3594..3698)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3779..3813
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     terminator      4557..4628
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4644..4671
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        4768..4843
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        4983..5058
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ttcagccaaa aaacttaaga ccgccggtct tgtccactac cttgcagtaa tgcggtggac
       61 aggatcggcg gttttctttt ctcttctcaa gaattcgcgg ccgcttctag agttatgaca
      121 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
      181 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
      241 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
      301 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
      361 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
      421 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
      481 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
      541 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
      601 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
      661 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
      721 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
      781 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
      841 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
      901 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
      961 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag
     1021 aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct
     1081 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag
     1141 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt
     1201 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc
     1261 ctacctgacg ctttttatcg caactctcta ctgtttctcc atataatacc cgtgtgaccc
     1321 gtgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     1381 gtggcaccga gtcggtgctt tttttgaagc ttgggcccga acaaaaactc atctcagaag
     1441 aggatctgaa tagcgccgtc gaccatcatc atcatcattg agtttaaacg gtctccagct
     1501 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
     1561 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     1621 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     1681 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1741 ttcgttttat ctgttgtttg tcggtgaact ggatccttac tcgagtctag actgcaggct
     1801 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1861 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccac
     1981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2401 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2701 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2761 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2821 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2881 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2941 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3001 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3061 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3121 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3181 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3241 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3301 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3361 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3421 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3481 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3541 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3601 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3661 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3721 cctgacgtct aagaaaccat tattatcatg acattaacca gttgataagt ccctaacttt
     3781 tacagctagc tcagtcctag gtattatgct agcctgaagt acgtctgagc gtgatacccg
     3841 ctcactgaag atggcccggt agggccgaaa cgtacctcta caaataattt tgtttaacta
     3901 tggactatgt ttgaaaggga gaaatactag atggcacgta ccccgagccg tagcagcatt
     3961 ggtagcctgc gtagtccgca tacccataaa gcaattctga ccagcaccat tgaaatcctg
     4021 aaagaatgtg gttatagcgg tctgagcatt gaaagcgttg cacgtcgtgc cggtgcaagc
     4081 aaaccgacca tttatcgttg gtggaccaat aaagcagcac tgattgccga agtgtatgaa
     4141 aatgaaagcg aacaggtgcg taaatttccg gatctgggta gctttaaagc cgatctggat
     4201 tttctgctgc gtaatctgtg gaaagtttgg cgtgaaacca tttgtggtga agcatttcgt
     4261 tgtgttattg cagaagcaca gctggaccct gcaaccctga cccagctgaa agatcagttt
     4321 atggaacgtc gtcgtgagat gccgaaaaaa ctggttgaaa atgccattag caatggtgaa
     4381 ctgccgaaag ataccaatcg tgaactgctg ctggatatga tttttggttt ttgttggtat
     4441 cgcctgctga ccgaacagct gaccgttgaa caggatattg aagaatttac cttcctgctg
     4501 attaatggtg tttgtccggg tacacagcgt taactagggc ccataccccc aggcatcaaa
     4561 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     4621 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatatac
     4681 gcgacgtacg gtggaatctg attcgttacc aattgacatg atacgaaacg taccgtatcg
     4741 ttaaggtata atacccgtgt gacccgtgtt ttagagctag aaatagcaag ttaaaataag
     4801 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctcggtacca
     4861 aagacgaaca ataagacgct gaaaagcgtc ttttttcgtt ttggtccttt agaattccag
     4921 aaatcatttt acaccaacgg gtcacacggg tattatgcta gccgccagct caatgaaaag
     4981 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     5041 gtggcaccga gtcggtgctt tttttctcgg taccaaattc cagaaaagag gcctcccgaa
     5101 aggggggcct tttttcgttt tggtccgctt
//
