LOCUS       sequence-461751-        4947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA-1 expression from an arabinose-inducible promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4947)
  AUTHORS   Pujar A, Pathania A, Hopper C, Pandi A, Calderon CR, Fugger M, Nowak
            T, Kushwaha M
  TITLE     Phage-mediated intercellular CRISPRi for biocomputation in bacterial
            consortia.
  JOURNAL   Nucleic Acids Res. 2024 Dec 27:gkae1256. doi: 10.1093/nar/gkae1256.
  PUBMED    39727169
REFERENCE   2  (bases 1 to 4947)
  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: "Nucleic 
            Acids Res. 2024 Dec 27:gkae1256. doi: 10.1093/nar/gkae1256."
COMMENT     Sequence Label: pSEVA19_AraC_pBAD-sgRNA-1
FEATURES             Location/Qualifiers
     source          1..4947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(14..100)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     115..137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             complement(169..1047)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1074..1358
                     /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     1281..1300
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_RNA        1380..1455
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      1520..1614
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1687..1778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1779..1847,1848..2639)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      2643..2682
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            2829..2937
                     /note="incP origin of transfer"
     rep_origin      2973..3561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3462..3481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3715..3732
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3747..3887
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3813..3832)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3973..3995
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3989..4180)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(4349..4858)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4544..4565)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4752..4771
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tctagggcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
       61 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctttaa ttaaagcgga
      121 taacaatttc acacaggagg ccgcctaggc cgcggccgcg cgatagaatt atgacaactt
      181 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
      241 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
      301 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
      361 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
      421 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
      481 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
      541 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
      601 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
      661 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
      721 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
      781 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
      841 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
      901 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
      961 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     1021 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac
     1081 caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct
     1141 aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat
     1201 gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt
     1261 gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag cggatcctac
     1321 ctgacgcttt ttatcgcaac tctctactgt ttctccataa tttccgtgcc tgggtatgag
     1381 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     1441 gcaccgagtc ggtgcttttc tgcaggcatg caagcttgcg gccgcgtcgt gactgggaaa
     1501 accctggcga ctagtcttgg actcctgttg atagatccag taatgacctc agaactccat
     1561 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaggg
     1621 gtccccaata attacgattt aaattagtag cccgcctaat gagcgggctt ttttttaatt
     1681 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1741 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagcattcag cattttcgtg
     1801 tggcgctgat tccgtttttt gcggcgtttt gcctgccggt gtttgcgcat ccggaaaccc
     1861 tggtgaaagt gaaagatgcg gaagatcaac tgggtgcgcg cgtgggctat attgaactgg
     1921 atctgaacag cggcaaaatt ctggaatctt ttcgtccgga agaacgtttt ccgatgatga
     1981 gcacctttaa agtgctgctg tgcggtgcgg ttctgagccg tgtggatgcg ggccaggaac
     2041 aactgggccg tcgtattcat tatagccaga acgatctggt ggaatatagc ccggtgaccg
     2101 aaaaacatct gaccgatggc atgaccgtgc gtgaactgtg cagcgcggcg attaccatga
     2161 gcgataacac cgcggcgaac ctgctgctga cgaccattgg cggtccgaaa gaactgaccg
     2221 cgtttctgca taacatgggc gatcatgtga cccgtctgga tcgttgggaa ccggaactga
     2281 acgaagcgat tccgaacgat gaacgtgata ccaccatgcc ggcagcaatg gcgaccaccc
     2341 tgcgtaaact gctgacgggt gagctgctga ccctggcaag ccgccagcaa ctgattgatt
     2401 ggatggaagc ggataaagtg gcgggtccgc tgctgcgtag cgcgctgccg gctggctggt
     2461 ttattgcgga taaaagcggt gcgggcgaac gtggcagccg tggcattatt gcggcgctgg
     2521 gcccggatgg taaaccgagc cgtattgtgg tgatttatac caccggcagc caggcgacga
     2581 tggatgaacg taaccgtcag attgcggaaa ttggcgcgag cctgattaaa cattggtaaa
     2641 ccgatacaat taaaggctcc ttttggagcc tttttttttg gacgaccctt gtccttttcc
     2701 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc
     2761 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg
     2821 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
     2881 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag
     2941 gccggccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3001 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3061 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3121 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3181 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3241 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3301 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3361 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3421 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3481 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3541 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3601 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3661 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3721 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3781 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3841 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     3901 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3961 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4021 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgcag cgattcacag atgtctgcct
     4081 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     4141 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     4201 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4261 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     4321 ggatgcggcg ggggcgcgag attgatttac cccggttgat aatcagaaaa gccccaaaaa
     4381 caggaagatt gtataagcaa atatttaaat tgtaagcgtt aatattttgt taaaattcgc
     4441 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4501 ttataaatca aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4561 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4621 tggcccacta cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc
     4681 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agcgaacgtg
     4741 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4801 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtaa
     4861 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4921 ttcgttccac tgaggcgcgc ccagctg
//
