LOCUS       sequence-159383-        6910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified helper phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6910)
  AUTHORS   Brodel AK, Jaramillo A, Isalan M
  TITLE     Engineering orthogonal dual transcription factors for multi-input 
            synthetic promoters.
  JOURNAL   Nat Commun. 2016 Dec 16;7:13858. doi: 10.1038/ncomms13858.
  PUBMED    27982027
REFERENCE   2  (bases 1 to 6910)
  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: "Nat 
            Commun."; date: "2016-12-16"; volume: "7:13858. doi"; pages: " 
            10.1038/ncomms13858"
COMMENT     Sequence Label: M13KO7-Delta-M13-Delta-geneIII-Delta-geneVI
FEATURES             Location/Qualifiers
     source          1..6910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             843..1106
                     /codon_start=1
                     /gene="V"
                     /product="pV"
                     /label=M13 gene V
                     /note="replication"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             1108..1209
                     /codon_start=1
                     /gene="VII"
                     /product="pVII"
                     /label=M13 gene VII
                     /note="minor coat protein"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             1206..1304
                     /codon_start=1
                     /gene="IX"
                     /product="pIX"
                     /label=M13 gene IX
                     /note="minor coat protein"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             1301..1522
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLALKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     terminator      1526..1574
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             1659..2705
                     /codon_start=1
                     /gene="I"
                     /product="pI"
                     /label=M13 gene I
                     /note="phage assembly"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     ITGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSLTYIDLCTVSIKKGN
                     SNEIVKCN"
     CDS             2683..3963
                     /codon_start=1
                     /gene="IV"
                     /product="pIV"
                     /label=M13 gene IV
                     /note="phage assembly"
                     /translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFVTWYSKQSGES
                     VIVSPDVKGTVTVYSSDVKPENLRNFFISVLRANNFDMVGSNPSIIQKYNPNNQDYIDE
                     LPSSDNQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLIRVVELFVKSNTSKSSNV
                     LSIDGSNLLVVSAPKDILDNLPQFLSTVDLPTDQILIEGLIFEVQQGDALDFSFAAGSQ
                     RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVRALKTNSHSKILSVPRILTLSGQ
                     KGSISVGQNVPFITGRVTGESANVNNPFQTIERQNVGISMSVFPVAMAGGNIVLDITSK
                     ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
                     LLFSSRSDSNEESTLYVLVKATIVRAL"
     primer_bind     complement(4153..4170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4287..4832)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4367..4386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5382..6197)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5382..6197)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6105..6124
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6189..6208
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(6512..6910,1..831)
                     /codon_start=1
                     /gene="II"
                     /product="pII"
                     /label=M13 gene II
                     /note="replication"
                     /translation="IDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSART
                     VEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNVF
                     GTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHLR
                     NVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQK
                     EQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIFD
                     LWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRLV
                     NEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRPD
                     WYNEPVLKIA"
ORIGIN
        1 aacgctacta ctattagtag aattgatgcc accttttcag ctcgcgcccc aaatgaaaat
       61 atagctaaac aggttattga ccatttgcga aatgtatcta atggtcaaac taaatctact
      121 cgttcgcaga attgggaatc aactgttaca tggaatgaaa cttccagaca ccgtacttta
      181 gttgcatatt taaaacatgt tgagctacag caccagattc agcaattaag ctctaagcca
      241 tccgcaaaaa tgacctctta tcaaaaggag caattaaagg tactctctaa tcctgacctg
      301 ttggagtttg cttccggtct ggttcgcttt gaagctcgaa ttaaaacgcg atatttgaag
      361 tctttcgggc ttcctcttaa tctttttgat gcaatccgct ttgcttctga ctataatagt
      421 cagggtaaag acctgatttt tgatttatgg tcattctcgt tttctgaact gtttaaagca
      481 tttgaggggg attcaatgaa tatttatgac gattccgcag tattggacgc tatccagtct
      541 aaacatttta ctattacccc ctctggcaaa acttcttttg caaaagcctc tcgctatttt
      601 ggtttttatc gtcgtctggt aaacgagggt tatgatagtg ttgctcttac tatgcctcgt
      661 aattcctttt ggcgttatgt atctgcatta gttgaatgtg gtattcctaa atctcaactg
      721 atgaatcttt ctacctgtaa taatgttgtt ccgttagttc gttttattaa cgtagatttt
      781 tcttcccaac gtcctgactg gtataatgag ccagttctta aaatcgcata aggtaattca
      841 caatgattaa agttgaaatt aaaccatctc aagcccaatt tactactcgt tctggtgttt
      901 ctcgtcaggg caagccttat tcactgaatg agcagctttg ttacgttgat ttgggtaatg
      961 aatatccggt tcttgtcaag attactcttg atgaaggtca gccagcctat gcgcctggtc
     1021 tgtacaccgt tcatctgtcc tctttcaaag ttggtcagtt cggttccctt atgattgacc
     1081 gtctgcgcct cgttccggct aagtaacatg gagcaggtcg cggatttcga cacaatttat
     1141 caggcgatga tacaaatctc cgttgtactt tgtttcgcgc ttggtataat cgctgggggt
     1201 caaagatgag tgttttagtg tattctttcg cctctttcgt tttaggttgg tgccttcgta
     1261 gtggcattac gtattttacc cgtttaatgg aaacttcctc atgaaaaagt ctttagccct
     1321 caaagcctct gtagccgttg ctaccctcgt tccgatgctg tctttcgctg ctgagggtga
     1381 cgatcccgca aaagcggcct ttaactccct gcaagcctca gcgaccgaat atatcggtta
     1441 tgcgtgggcg atggttgttg tcattgtcgg cgcaactatc ggtatcaagc tgtttaagaa
     1501 attcacctcg aaagcaagct gataaaccga tacaattaaa ggctcctttt ggagcctttt
     1561 tttttggaga ttttcaacgt gaaaaaatta ttattcgcaa ttcctttagt tgttcctttc
     1621 tattctcact ccgctgaaac tgttgaagga tcctaatcat ggctgtttat tttgtaactg
     1681 gcaaattagg ctctggaaag acgctcgtta gcgttggtaa gattcaggat aaaattgtag
     1741 ctgggtgcaa aatagcaact aatcttgatt taaggcttca aaacctcccg caagtcggga
     1801 ggttcgctaa aacgcctcgc gttcttagaa taccggataa gccttctata tctgatttgc
     1861 ttgctattgg gcgcggtaat gattcctacg atgaaaataa aaacggcttg cttgttctcg
     1921 atgagtgcgg tacttggttt aatacccgtt cttggaatga taaggaaaga cagccgatta
     1981 ttgattggtt tctacatgct cgtaaattag gatgggatat tatttttctt gttcaggact
     2041 tatctattgt tgataaacag gcgcgttctg cattagctga acatgttgtt tattgtcgtc
     2101 gtctggacag aattacttta ccttttgtcg gtactttata ttctcttatt actggctcga
     2161 aaatgcctct gcctaaatta catgttggcg ttgttaaata tggcgattct caattaagcc
     2221 ctactgttga gcgttggctt tatactggta agaatttgta taacgcatat gatactaaac
     2281 aggctttttc tagtaattat gattccggtg tttattctta tttaacgcct tatttatcac
     2341 acggtcggta tttcaaacca ttaaatttag gtcagaagat gaaattaact aaaatatatt
     2401 tgaaaaagtt ttctcgcgtt ctttgtcttg cgattggatt tgcatcagca tttacatata
     2461 gttatataac ccaacctaag ccggaggtta aaaaggtagt ctctcagacc tatgattttg
     2521 ataaattcac tattgactct tctcagcgtc ttaatctaag ctatcgctat gttttcaagg
     2581 attctaaggg aaaattaatt aatagcgacg atttacagaa gcaaggttat tcactcacat
     2641 atattgattt atgtactgtt tccattaaaa aaggtaattc aaatgaaatt gttaaatgta
     2701 attaattttg ttttcttgat gtttgtttca tcatcttctt ttgctcaggt aattgaaatg
     2761 aataattcgc ctctgcgcga ttttgtaact tggtattcaa agcaatcagg cgaatccgtt
     2821 attgtttctc ccgatgtaaa aggtactgtt actgtatatt catctgacgt taaacctgaa
     2881 aatctacgca atttctttat ttctgtttta cgtgcaaata attttgatat ggtaggttct
     2941 aacccttcca ttattcagaa gtataatcca aacaatcagg attatattga tgaattgcca
     3001 tcatctgata atcaggaata tgatgataat tccgctcctt ctggtggttt ctttgttccg
     3061 caaaatgata atgttactca aacttttaaa attaataacg ttcgggcaaa ggatttaata
     3121 cgagttgtcg aattgtttgt aaagtctaat acttctaaat cctcaaatgt attatctatt
     3181 gacggctcta atctattagt tgttagtgct cctaaagata ttttagataa ccttcctcaa
     3241 ttcctttcaa ctgttgattt gccaactgac cagatattga ttgagggttt gatatttgag
     3301 gttcagcaag gtgatgcttt agatttttca tttgctgctg gctctcagcg tggcactgtt
     3361 gcaggcggtg ttaatactga ccgcctcacc tctgttttat cttctgctgg tggttcgttc
     3421 ggtattttta atggcgatgt tttagggcta tcagttcgcg cattaaagac taatagccat
     3481 tcaaaaatat tgtctgtgcc acgtattctt acgctttcag gtcagaaggg ttctatctct
     3541 gttggccaga atgtcccttt tattactggt cgtgtgactg gtgaatctgc caatgtaaat
     3601 aatccatttc agacgattga gcgtcaaaat gtaggtattt ccatgagcgt ttttcctgtt
     3661 gcaatggctg gcggtaatat tgttctggat attaccagca aggccgatag tttgagttct
     3721 tctactcagg caagtgatgt tattactaat caaagaagta ttgctacaac ggttaatttg
     3781 cgtgatggac agactctttt actcggtggc ctcactgatt ataaaaacac ttctcaggat
     3841 tctggcgtac cgttcctgtc taaaatccct ttaatcggcc tcctgtttag ctcccgctct
     3901 gattctaacg aggaaagcac gttatacgtg ctcgtcaaag caaccatagt acgcgccctg
     3961 tagcgcaaaa aaccccgctt cggcggggtt ttttcgcacg gatcgcttca tgtggcacca
     4021 tagtacgcgc cctgtagcgc aaaaaacccc gcttcggcgg ggttttttcg cacggatcgc
     4081 ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat
     4141 atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga
     4201 aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga
     4261 agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac
     4321 gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg
     4381 tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc
     4441 attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc
     4501 agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt
     4561 atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc
     4621 agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa
     4681 ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt
     4741 tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc
     4801 aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaag gggtctgacg
     4861 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4921 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4981 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5041 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5101 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5161 atttatcagc aataaaccag ccagccgatt cgagctcgcc ccggggatcg accagttggt
     5221 gattttgaac ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg
     5281 atccttcaac tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg tcaagtcagc
     5341 gtaatgctct gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc
     5401 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     5461 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     5521 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     5581 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     5641 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     5701 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     5761 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     5821 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     5881 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     5941 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     6001 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     6061 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     6121 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc
     6181 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     6241 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg
     6301 tggctttccc cccccccccc tgcaggtctc gggctattct tttgatttat aagggatttt
     6361 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6421 taacaaaata ttaacgttta caatttaaat atttgcttat acaatcttcc tgtttttggg
     6481 gattttctga ttatcaaccg gggtacatgt gattgacatg ctagttttac gattaccgtt
     6541 catcgattct cttgtttgct ccagactctc aggcaatgac ctgatagcct ttgtagacct
     6601 ctcaaaaata gctaccctct ccggcatgaa tttatcagct agaacggttg aatatcatat
     6661 tgatggtgat ttgactgtct ccggcctttc tcaccctttt gaatctttac ctacacatta
     6721 ctcaggcatt gcatttaaaa tatatgaggg ttctaaaaat ttttatcctt gcgttgaaat
     6781 aaaggcttct cccgcaaaag tattacaggg tcataatgtt tttggtacaa ccgatttagc
     6841 tttatgctct gaggctttat tgcttaattt tgctaattct ttgccttgcc tgtatgattt
     6901 attggatgtt
//
