LOCUS       sequence-102060-        3838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of A2NT sgRNA from the arabinose-inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3838)
  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 3838)
  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-A2NT
FEATURES             Location/Qualifiers
     source          1..3838
                     /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             1421..1450
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1466..1483
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1539..1556)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1709..1755
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1808..1825)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1979..2567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2059..2078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2738..3529,3530..3598))
                     /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     3361..3380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3599..3703)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3771..3789
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3793..3836)
                     /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 ataataatac ccgtgtgacc
     1321 cgtgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     1381 agtggcaccg agtcggtgct tttttttgaa gcttgggccc gaacaaaaac tcatctcaga
     1441 agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt aaacggtctc
     1501 cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     1561 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     1621 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     1681 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     1741 ggcctttcgt tttatctgtt gtttgtcggt gaactggatc cttactcgag tctagactgc
     1801 aggcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1861 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1981 tccacaggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2401 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2521 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2581 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2641 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2701 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2761 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2821 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2881 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2941 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3001 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3061 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3121 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3181 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3241 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3301 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3361 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3421 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3481 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3541 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3601 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3661 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3721 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3781 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctg
//
