LOCUS       sequence-263408-        7175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Helper phage for M13 phage production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7175)
  AUTHORS   Brodel AK, Rodrigues R, Jaramillo A, Isalan M
  TITLE     Accelerated evolution of a minimal 63-amino acid dual transcription 
            factor.
  JOURNAL   Sci Adv. 2020 Jun 10;6(24):eaba2728. doi: 10.1126/sciadv.aba2728. 
            eCollection 2020 Jun.
  PUBMED    32577520
REFERENCE   2  (bases 1 to 7175)
  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: "Sci Adv."; 
            date: "2020-06-10"; volume: "6(24):eaba2728. doi"; pages: " 
            10.1126/sciadv.aba2728. eCollection 2020 Jun"
COMMENT     Sequence Label: M13KO7-Delta-PS-Delta-geneIII
FEATURES             Location/Qualifiers
     source          1..7175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(601..1416)
                     /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(601..1416)
                     /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     1324..1343
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1408..1427
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1731..2960
                     /codon_start=1
                     /gene="II"
                     /product="pII"
                     /label=M13 gene II
                     /note="replication"
                     /translation="IDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSART
                     VEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNVF
                     GTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHLR
                     NVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQK
                     EQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIFD
                     LWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRLV
                     NEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRPD
                     WYNEPVLKIA"
     CDS             2972..3235
                     /codon_start=1
                     /gene="V"
                     /product="pV"
                     /label=M13 gene V
                     /note="replication"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             3237..3338
                     /codon_start=1
                     /gene="VII"
                     /product="pVII"
                     /label=M13 gene VII
                     /note="minor coat protein"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             3335..3433
                     /codon_start=1
                     /gene="IX"
                     /product="pIX"
                     /label=M13 gene IX
                     /note="minor coat protein"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             3430..3651
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLALKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     terminator      3655..3703
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             3788..4126
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     CDS             4128..5174
                     /codon_start=1
                     /gene="I"
                     /product="pI"
                     /label=M13 gene I
                     /note="phage assembly"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     ITGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSLTYIDLCTVSIKKGN
                     SNEIVKCN"
     CDS             5152..6432
                     /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(6548..6565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6682..7175,1..52))
                     /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(6762..6781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca
       61 tcttattaag gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      121 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
      181 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
      241 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
      301 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
      361 cccacgctca ccggctccag atttatcagc aataaaccag ccagccgatt cgagctcgcc
      421 cggggatcga ccagttggtg attttgaact tttgctttgc cacggaacgg tctgcgttgt
      481 cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc
      541 gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga
      601 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
      661 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
      721 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
      781 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
      841 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
      901 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
      961 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     1021 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     1081 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     1141 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     1201 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     1261 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     1321 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     1381 cctcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     1441 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     1501 gagattttga gacacaacgt ggctttcccc ccccccccct gcaggtctcg ggctattctt
     1561 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1621 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaata tttgcttata
     1681 caatcttcct gtttttgggg attttctgat tatcaaccgg ggtacatgtg attgacatgc
     1741 tagttttacg attaccgttc atcgattctc ttgtttgctc cagactctca ggcaatgacc
     1801 tgatagcctt tgtagacctc tcaaaaatag ctaccctctc cggcatgaat ttatcagcta
     1861 gaacggttga atatcatatt gatggtgatt tgactgtctc cggcctttct cacccttttg
     1921 aatctttacc tacacattac tcaggcattg catttaaaat atatgagggt tctaaaaatt
     1981 tttatccttg cgttgaaata aaggcttctc ccgcaaaagt attacagggt cataatgttt
     2041 ttggtacaac cgatttagct ttatgctctg aggctttatt gcttaatttt gctaattctt
     2101 tgccttgcct gtatgattta ttggatgtta acgctactac tattagtaga attgatgcca
     2161 ccttttcagc tcgcgcccca aatgaaaata tagctaaaca ggttattgac catttgcgaa
     2221 atgtatctaa tggtcaaact aaatctactc gttcgcagaa ttgggaatca actgttacat
     2281 ggaatgaaac ttccagacac cgtactttag ttgcatattt aaaacatgtt gagctacagc
     2341 accagattca gcaattaagc tctaagccat ccgcaaaaat gacctcttat caaaaggagc
     2401 aattaaaggt actctctaat cctgacctgt tggagtttgc ttccggtctg gttcgctttg
     2461 aagctcgaat taaaacgcga tatttgaagt ctttcgggct tcctcttaat ctttttgatg
     2521 caatccgctt tgcttctgac tataatagtc agggtaaaga cctgattttt gatttatggt
     2581 cattctcgtt ttctgaactg tttaaagcat ttgaggggga ttcaatgaat atttatgacg
     2641 attccgcagt attggacgct atccagtcta aacattttac tattaccccc tctggcaaaa
     2701 cttcttttgc aaaagcctct cgctattttg gtttttatcg tcgtctggta aacgagggtt
     2761 atgatagtgt tgctcttact atgcctcgta attccttttg gcgttatgta tctgcattag
     2821 ttgaatgtgg tattcctaaa tctcaactga tgaatctttc tacctgtaat aatgttgttc
     2881 cgttagttcg ttttattaac gtagattttt cttcccaacg tcctgactgg tataatgagc
     2941 cagttcttaa aatcgcataa ggtaattcac aatgattaaa gttgaaatta aaccatctca
     3001 agcccaattt actactcgtt ctggtgtttc tcgtcagggc aagccttatt cactgaatga
     3061 gcagctttgt tacgttgatt tgggtaatga atatccggtt cttgtcaaga ttactcttga
     3121 tgaaggtcag ccagcctatg cgcctggtct gtacaccgtt catctgtcct ctttcaaagt
     3181 tggtcagttc ggttccctta tgattgaccg tctgcgcctc gttccggcta agtaacatgg
     3241 agcaggtcgc ggatttcgac acaatttatc aggcgatgat acaaatctcc gttgtacttt
     3301 gtttcgcgct tggtataatc gctgggggtc aaagatgagt gttttagtgt attctttcgc
     3361 ctctttcgtt ttaggttggt gccttcgtag tggcattacg tattttaccc gtttaatgga
     3421 aacttcctca tgaaaaagtc tttagccctc aaagcctctg tagccgttgc taccctcgtt
     3481 ccgatgctgt ctttcgctgc tgagggtgac gatcccgcaa aagcggcctt taactccctg
     3541 caagcctcag cgaccgaata tatcggttat gcgtgggcga tggttgttgt cattgtcggc
     3601 gcaactatcg gtatcaagct gtttaagaaa ttcacctcga aagcaagctg ataaaccgat
     3661 acaattaaag gctccttttg gagccttttt ttttggagat tttcaacgtg aaaaaattat
     3721 tattcgcaat tcctttagtt gttcctttct attctcactc cgctgaaact gttgaaggat
     3781 cctaatcatg ccagttcttt tgggtattcc gttattattg cgtttcctcg gtttccttct
     3841 ggtaactttg ttcggctatc tgcttacttt tcttaaaaag ggcttcggta agatagctat
     3901 tgctatttca ttgtttcttg ctcttattat tgggcttaac tcaattcttg tgggttatct
     3961 ctctgatatt agcgctcaat taccctctga ctttgttcag ggtgttcagt taattctccc
     4021 gtctaatgcg cttccctgtt tttatgttat tctctctgta aaggctgcta ttttcatttt
     4081 tgacgttaaa caaaaaatcg tttcttattt ggattgggat aaataatatg gctgtttatt
     4141 ttgtaactgg caaattaggc tctggaaaga cgctcgttag cgttggtaag attcaggata
     4201 aaattgtagc tgggtgcaaa atagcaacta atcttgattt aaggcttcaa aacctcccgc
     4261 aagtcgggag gttcgctaaa acgcctcgcg ttcttagaat accggataag ccttctatat
     4321 ctgatttgct tgctattggg cgcggtaatg attcctacga tgaaaataaa aacggcttgc
     4381 ttgttctcga tgagtgcggt acttggttta atacccgttc ttggaatgat aaggaaagac
     4441 agccgattat tgattggttt ctacatgctc gtaaattagg atgggatatt atttttcttg
     4501 ttcaggactt atctattgtt gataaacagg cgcgttctgc attagctgaa catgttgttt
     4561 attgtcgtcg tctggacaga attactttac cttttgtcgg tactttatat tctcttatta
     4621 ctggctcgaa aatgcctctg cctaaattac atgttggcgt tgttaaatat ggcgattctc
     4681 aattaagccc tactgttgag cgttggcttt atactggtaa gaatttgtat aacgcatatg
     4741 atactaaaca ggctttttct agtaattatg attccggtgt ttattcttat ttaacgcctt
     4801 atttatcaca cggtcggtat ttcaaaccat taaatttagg tcagaagatg aaattaacta
     4861 aaatatattt gaaaaagttt tctcgcgttc tttgtcttgc gattggattt gcatcagcat
     4921 ttacatatag ttatataacc caacctaagc cggaggttaa aaaggtagtc tctcagacct
     4981 atgattttga taaattcact attgactctt ctcagcgtct taatctaagc tatcgctatg
     5041 ttttcaagga ttctaaggga aaattaatta atagcgacga tttacagaag caaggttatt
     5101 cactcacata tattgattta tgtactgttt ccattaaaaa aggtaattca aatgaaattg
     5161 ttaaatgtaa ttaattttgt tttcttgatg tttgtttcat catcttcttt tgctcaggta
     5221 attgaaatga ataattcgcc tctgcgcgat tttgtaactt ggtattcaaa gcaatcaggc
     5281 gaatccgtta ttgtttctcc cgatgtaaaa ggtactgtta ctgtatattc atctgacgtt
     5341 aaacctgaaa atctacgcaa tttctttatt tctgttttac gtgcaaataa ttttgatatg
     5401 gtaggttcta acccttccat tattcagaag tataatccaa acaatcagga ttatattgat
     5461 gaattgccat catctgataa tcaggaatat gatgataatt ccgctccttc tggtggtttc
     5521 tttgttccgc aaaatgataa tgttactcaa acttttaaaa ttaataacgt tcgggcaaag
     5581 gatttaatac gagttgtcga attgtttgta aagtctaata cttctaaatc ctcaaatgta
     5641 ttatctattg acggctctaa tctattagtt gttagtgctc ctaaagatat tttagataac
     5701 cttcctcaat tcctttcaac tgttgatttg ccaactgacc agatattgat tgagggtttg
     5761 atatttgagg ttcagcaagg tgatgcttta gatttttcat ttgctgctgg ctctcagcgt
     5821 ggcactgttg caggcggtgt taatactgac cgcctcacct ctgttttatc ttctgctggt
     5881 ggttcgttcg gtatttttaa tggcgatgtt ttagggctat cagttcgcgc attaaagact
     5941 aatagccatt caaaaatatt gtctgtgcca cgtattctta cgctttcagg tcagaagggt
     6001 tctatctctg ttggccagaa tgtccctttt attactggtc gtgtgactgg tgaatctgcc
     6061 aatgtaaata atccatttca gacgattgag cgtcaaaatg taggtatttc catgagcgtt
     6121 tttcctgttg caatggctgg cggtaatatt gttctggata ttaccagcaa ggccgatagt
     6181 ttgagttctt ctactcaggc aagtgatgtt attactaatc aaagaagtat tgctacaacg
     6241 gttaatttgc gtgatggaca gactctttta ctcggtggcc tcactgatta taaaaacact
     6301 tctcaggatt ctggcgtacc gttcctgtct aaaatccctt taatcggcct cctgtttagc
     6361 tcccgctctg attctaacga ggaaagcacg ttatacgtgc tcgtcaaagc aaccatagta
     6421 cgcgccctgt agcgcaaaaa accccgcttc ggcggggttt tttcgcacgg atcgcttcat
     6481 gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt
     6541 ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg
     6601 cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga
     6661 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
     6721 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6781 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     6841 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     6901 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     6961 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     7021 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     7081 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     7141 gctcagagaa ccttcgaaaa accgccctgc aaggc
//
