LOCUS       sequence-372770-        5624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M13 packaging signal, LacZ-alpha gene, and M13 gIII gene (also 
            called gp3, or g3p) cloned into the pSB1K3 vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5624)
  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 5624)
  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: pSB1K3_M13ps_LacZ-alpha_gIII
FEATURES             Location/Qualifiers
     source          1..5624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(213..234)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     421..440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     729..748
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     misc_feature    846..866
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     protein_bind    complement(877..910)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(1039..2313)
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     terminator      complement(2318..2366)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     protein_bind    2442..2463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2478..2483,2484..2501,2502..2508)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2516..2532
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2521..2543
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2540..2556
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2540..2556
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2552..3088
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFAS
                     WRNSEEARTDRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHG
                     YDAPIYTNVTYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVPIGKSKT
                     DFPFIIFEIYFLNSL"
     primer_bind     complement(2599..2615)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2608..2630)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3385..3401)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3500..3517)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3671..4259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3751..3770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4381..5190)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(5191..5282)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5349..5368
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    5575..5608
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    join(5619..5624,1..14)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
ORIGIN
        1 gcggccgctt ctagggaacc ccggttgata atcagaaaag ccccaaaaac aggaagattg
       61 tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
      121 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      181 aagaatagcc cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
      241 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
      301 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
      361 accctaaagg gagcccccga tttagagctt gacggggaaa gcgaacgtgg cgagaaagga
      421 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
      481 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggccgggatc tcgacgctct
      541 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
      601 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
      661 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
      721 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
      781 ccggccacga tgcgtccggc gtagaggatc gagatcgatc tcgatcccgc gaaattaata
      841 cgatgtacta gtagcggccg ctgcagaaaa tgaagtgaag ttcctatact ttctagagaa
      901 taggaacttc tatagtgagt cgaataaggg cgacacaaaa tttattctaa atgcataata
      961 aatactgata acatcttata gtttgtatta tattttgtat tatcgttgac atgtataatt
     1021 ttgattatcc ctgcaccgtt aagactcctt attacgcagt atgttagcaa acgtagaaaa
     1081 tacatacata aaggtggcaa catataaaag aaacgcaaag acaccacgga ataagtttat
     1141 tttgtcacaa tcaatagaaa attcatatgg tttaccagcg ccaaagacaa aagggcgaca
     1201 ttcaaccgat tgagggaggg aaggtaaata ttgacggaaa ttattcatta aaggtgaatt
     1261 atcaccgtca ccgacttgag ccatttggga attagagcca gcaaaatcac cagtagcacc
     1321 attaccatta gcaaggccgg aaacgtcacc aatgaaacca tcgatagcag caccgtaatc
     1381 agtagcgaca gaatcaagtt tgcctttagc gtcagactgt agcgcgtttt catcggcatt
     1441 ttcggtcata gcccccttat tagcgtttgc catcttttca taatcaaaat caccggaacc
     1501 agagccacca ccggaaccgc ctccctcaga gccgccaccc tcagaaccgc caccctcaga
     1561 gccaccaccc tcagagccgc caccagaacc accaccagag ccgccgccag cattgacagg
     1621 aggttgaggc aggtcagacg attggccttg atattcacaa acgaatggat cctcattaaa
     1681 gccagaatgg aaagcgcagt ctctgaattt accgttccag taagcgtcat acatggcttt
     1741 tgatgataca ggagtgtact ggtaataagt tttaacgggg tcagtgcctt gagtaacagt
     1801 gcccgtataa acagttaatg ccccctgcct atttcggaac ctattattct gaaacatgaa
     1861 agtattaaga ggctgagact cctcaagaga aggattagga ttagcggggt tttgctcagt
     1921 accaggcgga taagtgccgt cgagagggtt gatataagta tagcccggaa taggtgtatc
     1981 accgtactca ggaggtttag taccgccacc ctcagaaccg ccaccctcag aaccgccacc
     2041 ctcagagcca ccaccctcat tttcagggat agcaagccca ataggaaccc atgtaccgta
     2101 acactgagtt tcgtcaccag tacaaactac aacgcctgta gcattccaca gacagccctc
     2161 atagttagcg taacgatcta aagttttgtc gtctttccag acgttagtaa atgaattttc
     2221 tgtatggggt tttgctaaac aactttcaac agtttcagcg gagtgagaat agaaaggaac
     2281 aactaaagga attgcgaata ataatttttt cacgttgaaa atctccaaaa aaaaaggctc
     2341 caaaaggagc ctttaattgt atcggtttat cagcttgctt tcgaggtgaa tttcttaaac
     2401 agcttgacga cagggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2461 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2521 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc
     2581 ctgcaggtcc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2641 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2701 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
     2761 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga
     2821 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
     2881 caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
     2941 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
     3001 aattattttt gatggcgttc ctattggtaa atcaaaaact gattttccct ttattatttt
     3061 cgagatttat tttcttaatt ctctttaaca aactagaaat attgtatata caaaaaatca
     3121 taaataatag atgaatagtt taattatagg tgttcatcaa tcgaaaaagc aacgtatctt
     3181 atttaaagtg cgttgctttt ttctcattta taaggttaaa taattctcat atatcaagca
     3241 aagtgacagg cgcccttaaa tattctgaca aatgctcttt ccctaaactc cccccataaa
     3301 aaaacccgcc gaagcgggtt tttacgttat ttgcggatta acgattactc gttatcagaa
     3361 ccgcccaggg ggcccgagct taagactggc cgtcgtttta caacacagaa agagtttgta
     3421 gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag tttgatgcct ggcagttccc
     3481 tactctcgcc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3541 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3601 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3661 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3721 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3781 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3841 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3901 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3961 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4021 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4081 agtggtgggc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4141 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4201 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4261 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg cgcgtaactc
     4321 acgttaaggg attttggtca tgagcttgcg ccgtcccgtc aagtcagcgt aatgctctgc
     4381 ttagaaaaac tcatcgagca tcaaatgaaa ttgcaattta ttcatatcag gattatcaat
     4441 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     4501 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     4561 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     4621 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     4681 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     4741 cgcctgagcg aggcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     4801 gtgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     4861 ttcttctaat acctggaacg ctgtttttcc ggggatcgca gtggtgagta accatgcatc
     4921 atcaggagta cggataaaat gcttgatggt cggaagtggc ataaattccg tcagccagtt
     4981 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     5041 caactctggc gcatcgggct tcccatacaa gcgatagatt gtcgcacctg attgcccgac
     5101 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     5161 cctcgacgtt tcccgttgaa tatggctcat attcttcctt tttcaatatt attgaagcat
     5221 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5281 aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga gcccatttat
     5341 acctgaatat ggctcataac accccttgtt tgcctggcgg cagtagcgcg gtggtcccac
     5401 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggactc
     5461 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     5521 tgggcctttc gcccgggcta attagggggt gtcgccctta ttcgactcta tagtgaagtt
     5581 cctattctct agaaagtata ggaacttctg aagtggggga attc
//
