LOCUS       sequence-377863-        7768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M13 helper genes cloned into the pSEVA631 vector, except the minor 
            coat protein coding gene gIII (also called gp3, or g3p). This 
            plasmid does not carry an M13 packaging signal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7768)
  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 7768)
  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: pSEVA63_Helper-Delta-gIII
FEATURES             Location/Qualifiers
     source          1..7768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..23
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             173..1405
                     /codon_start=1
                     /gene="II"
                     /product="pII"
                     /label=M13 gene II
                     /note="replication"
                     /translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
                     TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
                     FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
                     RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
                     KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
                     DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
                     VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
                     DWYNEPVLKIA"
     CDS             1417..1680
                     /codon_start=1
                     /gene="V"
                     /product="pV"
                     /label=M13 gene V
                     /note="replication"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             1682..1783
                     /codon_start=1
                     /gene="VII"
                     /product="pVII"
                     /label=M13 gene VII
                     /note="minor coat protein"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             1780..1878
                     /codon_start=1
                     /gene="IX"
                     /product="pIX"
                     /label=M13 gene IX
                     /note="minor coat protein"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             1875..2096
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLALKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     terminator      2100..2148
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             2197..2535
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     CDS             2537..3583
                     /codon_start=1
                     /gene="I"
                     /product="pI"
                     /label=M13 gene I
                     /note="phage assembly"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     ITGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSLTYIDLCTVSIKKGN
                     SNEIVKCN"
     CDS             3561..4841
                     /codon_start=1
                     /gene="IV"
                     /product="pIV"
                     /label=M13 gene IV
                     /note="phage assembly"
                     /translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFVTWYSKQSGES
                     VIVSPDVKGTVTVYSSDVKPENLRNFFISVLRANNFDMVGSNPSIIQKYNPNNQDYIDE
                     LPSSDNQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLIRVVELFVKSNTSKSSNV
                     LSIDGSNLLVVSAPKDILDNLPQFLSTVDLPTDQILIEGLIFEVQQGDALDFSFAAGSQ
                     RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVRALKTNSHSKILSVPRILTLSGQ
                     KGSISVGQNVPFITGRVTGESANVNNPFQTIERQNVGISMSVFPVAMAGGNIVLDITSK
                     ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
                     LLFSSRSDSNEESTLYVLVKATIVRAL"
     terminator      4914..5008
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(5039..5044,5045..5061,5062..5067)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5256..5789
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            6000..6108
                     /note="incP origin of transfer"
     CDS             complement(6119..6781)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      6782..7553
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(7668..7754)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcggataac aatttcacac aggaggccgc ctaggcctat tggttaaaaa atgagctgat
       61 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt aaatatttgc
      121 ttatacaatc ttcctgtttt tggggctttt ctgattatca accggggtac atatgattga
      181 catgctagtt ttacgattac cgttcatcga ttctcttgtt tgctccagac tctcaggcaa
      241 tgacctgata gcctttgtag acctctcaaa aatagctacc ctctccggca tgaatttatc
      301 agctagaacg gttgaatatc atattgatgg tgatttgact gtctccggcc tttctcaccc
      361 ttttgaatct ttacctacac attactcagg cattgcattt aaaatatatg agggttctaa
      421 aaatttttat ccttgcgttg aaataaaggc ttctcccgca aaagtattac agggtcataa
      481 tgtttttggt acaaccgatt tagctttatg ctctgaggct ttattgctta attttgctaa
      541 ttctttgcct tgcctgtatg atttattgga tgttaacgct actactatta gtagaattga
      601 tgccaccttt tcagctcgcg ccccaaatga aaatatagct aaacaggtta ttgaccattt
      661 gcgaaatgta tctaatggtc aaactaaatc tactcgttcg cagaattggg aatcaactgt
      721 tacatggaat gaaacttcca gacaccgtac tttagttgca tatttaaaac atgttgagct
      781 acagcaccag attcagcaat taagctctaa gccatccgca aaaatgacct cttatcaaaa
      841 ggagcaatta aaggtactct ctaatcctga cctgttggag tttgcttccg gtctggttcg
      901 ctttgaagct cgaattaaaa cgcgatattt gaagtctttc gggcttcctc ttaatctttt
      961 tgatgcaatc cgctttgctt ctgactataa tagtcagggt aaagacctga tttttgattt
     1021 atggtcattc tcgttttctg aactgtttaa agcatttgag ggggattcaa tgaatattta
     1081 tgacgattcc gcagtattgg acgctatcca gtctaaacat tttactatta ccccctctgg
     1141 caaaacttct tttgcaaaag cctctcgcta ttttggtttt tatcgtcgtc tggtaaacga
     1201 gggttatgat agtgttgctc ttactatgcc tcgtaattcc ttttggcgtt atgtatctgc
     1261 attagttgaa tgtggtattc ctaaatctca actgatgaat ctttctacct gtaataatgt
     1321 tgttccgtta gttcgtttta ttaacgtaga tttttcttcc caacgtcctg actggtataa
     1381 tgagccagtt cttaaaatcg cataaggtaa ttcacaatga ttaaagttga aattaaacca
     1441 tctcaagccc aatttactac tcgttctggt gtttctcgtc agggcaagcc ttattcactg
     1501 aatgagcagc tttgttacgt tgatttgggt aatgaatatc cggttcttgt caagattact
     1561 cttgatgaag gtcagccagc ctatgcgcct ggtctgtaca ccgttcatct gtcctctttc
     1621 aaagttggtc agttcggttc ccttatgatt gaccgtctgc gcctcgttcc ggctaagtaa
     1681 catggagcag gtcgcggatt tcgacacaat ttatcaggcg atgatacaaa tctccgttgt
     1741 actttgtttc gcgcttggta taatcgctgg gggtcaaaga tgagtgtttt agtgtattct
     1801 ttcgcctctt tcgttttagg ttggtgcctt cgtagtggca ttacgtattt tacccgttta
     1861 atggaaactt cctcatgaaa aagtctttag ccctcaaagc ctctgtagcc gttgctaccc
     1921 tcgttccgat gctgtctttc gctgctgagg gtgacgatcc cgcaaaagcg gcctttaact
     1981 ccctgcaagc ctcagcgacc gaatatatcg gttatgcgtg ggcgatggtt gttgtcattg
     2041 tcggcgcaac tatcggtatc aagctgttta agaaattcac ctcgaaagca agctgataaa
     2101 ccgatacaat taaaggctcc ttttggagcc tttttttttg gagattttca accgacctac
     2161 gtttgctaac atactgcgta ataaggagtc ttaatcatgc cagttctttt gggtattccg
     2221 ttattattgc gtttcctcgg tttccttctg gtaactttgt tcggctatct gcttactttt
     2281 cttaaaaagg gcttcggtaa gatagctatt gctatttcat tgtttcttgc tcttattatt
     2341 gggcttaact caattcttgt gggttatctc tctgatatta gcgctcaatt accctctgac
     2401 tttgttcagg gtgttcagtt aattctcccg tctaatgcgc ttccctgttt ttatgttatt
     2461 ctctctgtaa aggctgctat tttcattttt gacgttaaac aaaaaatcgt ttcttatttg
     2521 gattgggata aataatatgg ctgtttattt tgtaactggc aaattaggct ctggaaagac
     2581 gctcgttagc gttggtaaga ttcaggataa aattgtagct gggtgcaaaa tagcaactaa
     2641 tcttgattta aggcttcaaa acctcccgca agtcgggagg ttcgctaaaa cgcctcgcgt
     2701 tcttagaata ccggataagc cttctatatc tgatttgctt gctattgggc gcggtaatga
     2761 ttcctacgat gaaaataaaa acggcttgct tgttctcgat gagtgcggta cttggtttaa
     2821 tacccgttct tggaatgata aggaaagaca gccgattatt gattggtttc tacatgctcg
     2881 taaattagga tgggatatta tttttcttgt tcaggactta tctattgttg ataaacaggc
     2941 gcgttctgca ttagctgaac atgttgttta ttgtcgtcgt ctggacagaa ttactttacc
     3001 ttttgtcggt actttatatt ctcttattac tggctcgaaa atgcctctgc ctaaattaca
     3061 tgttggcgtt gttaaatatg gcgattctca attaagccct actgttgagc gttggcttta
     3121 tactggtaag aatttgtata acgcatatga tactaaacag gctttttcta gtaattatga
     3181 ttccggtgtt tattcttatt taacgcctta tttatcacac ggtcggtatt tcaaaccatt
     3241 aaatttaggt cagaagatga aattaactaa aatatatttg aaaaagtttt ctcgcgttct
     3301 ttgtcttgcg attggatttg catcagcatt tacatatagt tatataaccc aacctaagcc
     3361 ggaggttaaa aaggtagtct ctcagaccta tgattttgat aaattcacta ttgactcttc
     3421 tcagcgtctt aatctaagct atcgctatgt tttcaaggat tctaagggaa aattaattaa
     3481 tagcgacgat ttacagaagc aaggttattc actcacatat attgatttat gtactgtttc
     3541 cattaaaaaa ggtaattcaa atgaaattgt taaatgtaat taattttgtt ttcttgatgt
     3601 ttgtttcatc atcttctttt gctcaggtaa ttgaaatgaa taattcgcct ctgcgcgatt
     3661 ttgtaacttg gtattcaaag caatcaggcg aatccgttat tgtttctccc gatgtaaaag
     3721 gtactgttac tgtatattca tctgacgtta aacctgaaaa tctacgcaat ttctttattt
     3781 ctgttttacg tgcaaataat tttgatatgg taggttctaa cccttccatt attcagaagt
     3841 ataatccaaa caatcaggat tatattgatg aattgccatc atctgataat caggaatatg
     3901 atgataattc cgctccttct ggtggtttct ttgttccgca aaatgataat gttactcaaa
     3961 cttttaaaat taataacgtt cgggcaaagg atttaatacg agttgtcgaa ttgtttgtaa
     4021 agtctaatac ttctaaatcc tcaaatgtat tatctattga cggctctaat ctattagttg
     4081 ttagtgctcc taaagatatt ttagataacc ttcctcaatt cctttcaact gttgatttgc
     4141 caactgacca gatattgatt gagggtttga tatttgaggt tcagcaaggt gatgctttag
     4201 atttttcatt tgctgctggc tctcagcgtg gcactgttgc aggcggtgtt aatactgacc
     4261 gcctcacctc tgttttatct tctgctggtg gttcgttcgg tatttttaat ggcgatgttt
     4321 tagggctatc agttcgcgca ttaaagacta atagccattc aaaaatattg tctgtgccac
     4381 gtattcttac gctttcaggt cagaagggtt ctatctctgt tggccagaat gtccctttta
     4441 ttactggtcg tgtgactggt gaatctgcca atgtaaataa tccatttcag acgattgagc
     4501 gtcaaaatgt aggtatttcc atgagcgttt ttcctgttgc aatggctggc ggtaatattg
     4561 ttctggatat taccagcaag gccgatagtt tgagttcttc tactcaggca agtgatgtta
     4621 ttactaatca aagaagtatt gctacaacgg ttaatttgcg tgatggacag actcttttac
     4681 tcggtggcct cactgattat aaaaacactt ctcaggattc tggcgtaccg ttcctgtcta
     4741 aaatcccttt aatcggcctc ctgtttagct cccgctctga ttctaacgag gaaagcacgt
     4801 tatacgtgct cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4861 catgcaagct tgcggccgcg tcgtgactgg gaaaaccctg gcgactagtc ttggactcct
     4921 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     4981 ccgccgggcg ttttttattg gtgagaatcc aggggtcccc aataattacg atttaaattt
     5041 gacataagcc tgttcggttc gtaaactgta atgcaagtag cgtatgcgct cacgcaactg
     5101 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     5161 ttatgactgt ttttttgtac agcctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     5221 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagc aacgatgtta
     5281 cgcagcaggg cagtcgccct aaaacaaagt taggtggctc aagtatgggc atcattcgca
     5341 catgtaggct cggccctgac caagtcaaat ccatgcgggc tgctcttgat cttttcggtc
     5401 gtgagttcgg agacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga
     5461 acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg
     5521 gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag atctatatct
     5581 atgatctcgc agtctccgga gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc
     5641 tcctcaagca tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg
     5701 gtgacgatcc cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact
     5761 ttgatatcga cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg
     5821 ccgcggagtt gttcggtaaa ttggacaacg gtccgcgcgt tgtccttttc cgctgcataa
     5881 ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat
     5941 acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg
     6001 ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat
     6061 tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccgta ggccggccct
     6121 accggcgcgg cagcgttacc cgtgtcggcg gctccaacgg ctcgccatcg tccagaaaac
     6181 acggctcatc gggcatcggc aggcgctgct gcccgcgccg ttcccattcc tccgtttcgg
     6241 tcaaggctgg caggtctggt tccatgcccg gaatgccggg ctggctgggc ggctcctcgc
     6301 cggggccggt cggtagttgc tgctcgcccg gatacagggt cgggatgcgg cgcaggtcgc
     6361 catgccccaa cagcgattcg tcctggtcgt cgtgatcaac caccacggcg gcactgaaca
     6421 ccgacaggcg caactggtcg cggggctggc cccacgccac gcggtcattg accacgtagg
     6481 ccgacacggt gccggggccg ttgagcttca cgacggagat ccagcgctcg gccaccaagt
     6541 ccttgactgc gtattggacc gtccgcaaag aacgtccgat gagcttggaa agtgtcttct
     6601 ggctgaccac cacggcgttc tggtggccca tctgcgccac gaggtgatgc agcagcattg
     6661 ccgccgtggg tttcctcgca ataagcccgg cccacgcctc atgcgctttg cgttccgttt
     6721 gcacccagtg accgggcttg ttcttggctt gaatgccgat ttctctggac tgcgtggcca
     6781 tgcttatctc catgcggtag gggtgccgca cggttgcggc accatgcgca atcagctgca
     6841 acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt
     6901 ttctttaacc tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc
     6961 ccttttttaa gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg
     7021 tgcccaaaga agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg
     7081 gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag
     7141 gtggcgctgc ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg
     7201 cgggcttcgc ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa
     7261 ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac
     7321 cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa
     7381 ttgatggggc aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg
     7441 ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc
     7501 ttgtccagca gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc
     7561 catcgtccac atatccacgg gctggcaagg gagcgcagcg accgcgcagg gcgaagcccg
     7621 gagagcaagc ccgtaggggg ggcgcgccca gctgtctagg gcggcggatt tgtcctactc
     7681 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc
     7741 tttcgtttta tttgatgcct ttaattaa
//
