LOCUS       sequence-102133-        3837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of A2T sgRNA from the arabinose-inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3837)
  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 3837)
  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-PBAD-sgRNA-A2T
FEATURES             Location/Qualifiers
     source          1..3837
                     /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        1324..1399
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1420..1449
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1465..1482
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1538..1555)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1708..1754
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1807..1824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1978..2566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2058..2077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2737..3528,3529..3597))
                     /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     3360..3379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3598..3702)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3770..3788
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3792..3835)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
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 atatttacac caacgggtca
     1321 cacgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     1381 agtggcaccg agtcggtgct ttttttgaag cttgggcccg aacaaaaact catctcagaa
     1441 gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta aacggtctcc
     1501 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
     1561 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
     1621 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
     1681 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1741 gcctttcgtt ttatctgttg tttgtcggtg aactggatcc ttactcgagt ctagactgca
     1801 ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1861 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1921 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1981 ccacaggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2041 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2101 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2161 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2221 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2281 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2341 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2401 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2461 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2521 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2581 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2641 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2701 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2761 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2821 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2881 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2941 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3001 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3061 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3121 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3181 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3241 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3301 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3361 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3421 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3481 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3541 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3601 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3661 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3721 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3781 cacgaggcag aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttctg
//
