LOCUS       sequence-424894-        6864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Helper plasmid that allows phagemid production upon complementation 
            with a phagemid expressing pI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6864)
  AUTHORS   Gretton D, Wang B, Edison R, Foner L, Berlips J, Vogel T, Kysel M, 
            Chen W, Sage-Ling F, Hauwe LV, Wooster S, Weinstein-Raun B, 
            DeBenedictis EA, Liu AB, Chory E, Cui H, Li X, Dong J, Fabrega A, 
            Dennison C, Don O, Ye CT, Uberoy K, Rivest RL, Gao M, Yu Y, Baum C, 
            Damgard I, Yao AC, Esvelt KM
  TITLE     Random adversarial threshold search enables automated DNA screening
  JOURNAL   bioRxiv 2024.03.20.585782
REFERENCE   2  (bases 1 to 6864)
  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: "bioRxiv 
            2024.03.20.585782"
COMMENT     Sequence Label: M13cp-dg1 hp
FEATURES             Location/Qualifiers
     source          1..6864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             31..252
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     terminator      256..303
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             308..1582
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKSHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDLFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     CDS             1585..1923
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     CDS             1925..3205
                     /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(3309..3326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3443..3988)
                     /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(3523..3542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4179..4838)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4753..4772
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4839..4941)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5193..6425
                     /codon_start=1
                     /gene="II"
                     /product="pII"
                     /label=M13 gene II
                     /note="replication"
                     /translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGINLSAR
                     TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
                     FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
                     RNVSNGQTKSTRSQNWESTVIWNETSRHRTLVAYLKHVELQHYIQQLSSKPSAKMTSYQ
                     KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
                     DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
                     VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
                     DWYNEPVLKIA"
     CDS             6437..6700
                     /codon_start=1
                     /gene="V"
                     /product="pV"
                     /label=M13 gene V
                     /note="replication"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             6702..6803
                     /codon_start=1
                     /gene="VII"
                     /product="pVII"
                     /label=M13 gene VII
                     /note="minor coat protein"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             join(6800..6864,1..34)
                     /codon_start=1
                     /gene="IX"
                     /product="pIX"
                     /label=M13 gene IX
                     /note="minor coat protein"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
ORIGIN
        1 gtattttacc cgtttaatgg aaacttcctc atgaaaaagt ctttagtcct caaagcctct
       61 gtagccgttg ctaccctcgt tccgatgctg tctttcgctg ctgagggtga cgatcccgca
      121 aaagcggcct ttaactccct gcaagcctca gcgaccgaat atatcggtta tgcgtgggcg
      181 atggttgttg tcattgtcgg cgcaactatc ggtatcaagc tgtttaagaa attcacctcg
      241 aaagcaagct gataaaccga tacaattaaa ggctcctttt ggagcctttt ttttggagat
      301 tttcaacgtg aaaaaattat tattcgcaat tcctttagtt gttcctttct attctcactc
      361 cgctgaaact gttgaaagtt gtttagcaaa atcccataca gaaaattcat ttactaacgt
      421 ctggaaagac gacaaaactt tagatcgtta cgctaactat gagggctgtc tgtggaatgc
      481 tacaggcgtt gtagtttgta ctggtgacga aactcagtgt tacggtacat gggttcctat
      541 tgggcttgct atccctgaaa atgagggtgg tggctctgag ggtggcggtt ctgagggtgg
      601 cggttctgag ggtggcggta ctaaacctcc tgagtacggt gatacaccta ttccgggcta
      661 tacttatatc aaccctctcg acggcactta tccgcctggt actgagcaaa accccgctaa
      721 tcctaatcct tctcttgagg agtctcagcc tcttaatact ttcatgtttc agaataatag
      781 gttccgaaat aggcaggggg cattaactgt ttatacgggc actgttactc aaggcactga
      841 ccccgttaaa acttattacc agtacactcc tgtatcatca aaagccatgt atgacgctta
      901 ctggaacggt aaattcagag actgcgcttt ccattctggc tttaatgagg atttatttgt
      961 ttgtgaatat caaggccaat cgtctgacct gcctcaacct cctgtcaatg ctggcggcgg
     1021 ctctggtggt ggttctggtg gcggctctga gggtggtggc tctgagggtg gcggttctga
     1081 gggtggcggc tctgagggag gcggttccgg tggtggctct ggttccggtg attttgatta
     1141 tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa aatgccgatg aaaacgcgct
     1201 acagtctgac gctaaaggca aacttgattc tgtcgctact gattacggtg ctgctatcga
     1261 tggtttcatt ggtgacgttt ccggccttgc taatggtaat ggtgctactg gtgattttgc
     1321 tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat aattcacctt taatgaataa
     1381 tttccgtcaa tatttacctt ccctccctca atcggttgaa tgtcgccctt ttgtctttgg
     1441 cgctggtaaa ccatatgaat tttctattga ttgtgacaaa ataaacttat tccgtggtgt
     1501 ctttgcgttt cttttatatg ttgccacctt tatgtatgta ttttctacgt ttgctaacat
     1561 actgcgtaat aaggagtctt aatcatgcca gttcttttgg gtattccgtt attattgcgt
     1621 ttcctcggtt tccttctggt aactttgttc ggctatctgc ttacttttct taaaaagggc
     1681 ttcggtaaga tagctattgc tatttcattg tttcttgctc ttattattgg gcttaactca
     1741 attcttgtgg gttatctctc tgatattagc gctcaattac cctctgactt tgttcagggt
     1801 gttcagttaa ttctcccgtc taatgcgctt ccctgttttt atgttattct ctctgtaaag
     1861 gctgctattt tcatttttga cgttaaacaa aaaatcgttt cttatttgga ttgggataaa
     1921 taatatgaaa ttgttaaatg taattaattt tgttttcttg atgtttgttt catcatcttc
     1981 ttttgctcag gtaattgaaa tgaataattc gcctctgcgc gattttgtaa cttggtattc
     2041 aaagcaatca ggcgaatccg ttattgtttc tcccgatgta aaaggtactg ttactgtata
     2101 ttcatctgac gttaaacctg aaaatctacg caatttcttt atttctgttt tacgtgcaaa
     2161 taattttgat atggtaggtt ctaacccttc cattattcag aagtataatc caaacaatca
     2221 ggattatatt gatgaattgc catcatctga taatcaggaa tatgatgata attccgctcc
     2281 ttctggtggt ttctttgttc cgcaaaatga taatgttact caaactttta aaattaataa
     2341 cgttcgggca aaggatttaa tacgagttgt cgaattgttt gtaaagtcta atacttctaa
     2401 atcctcaaat gtattatcta ttgacggctc taatctatta gttgttagtg ctcctaaaga
     2461 tattttagat aaccttcctc aattcctttc aactgttgat ttgccaactg accagatatt
     2521 gattgagggt ttgatatttg aggttcagca aggtgatgct ttagattttt catttgctgc
     2581 tggctctcag cgtggcactg ttgcaggcgg tgttaatact gaccgcctca cctctgtttt
     2641 atcttctgct ggtggttcgt tcggtatttt taatggcgat gttttagggc tatcagttcg
     2701 cgcattaaag actaatagcc attcaaaaat attgtctgtg ccacgtattc ttacgctttc
     2761 aggtcagaag ggttctatct ctgttggcca gaatgtccct tttattactg gtcgtgtgac
     2821 tggtgaatct gccaatgtaa ataatccatt tcagacgatt gagcgtcaaa atgtaggtat
     2881 ttccatgagc gtttttcctg ttgcaatggc tggcggtaat attgttctgg atattaccag
     2941 caaggccgat agtttgagtt cttctactca ggcaagtgat gttattacta atcaaagaag
     3001 tattgctaca acggttaatt tgcgtgatgg acagactctt ttactcggtg gcctcactga
     3061 ttataaaaac acttctcagg attctggcgt accgttcctg tctaaaatcc ctttaatcgg
     3121 cctcctgttt agctcccgct ctgattctaa cgaggaaagc acgttatacg tgctcgtcaa
     3181 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ctgcagagaa catggcttca
     3241 tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat
     3301 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg
     3361 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     3421 agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa
     3481 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3541 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     3601 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
     3661 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
     3721 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
     3781 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
     3841 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
     3901 agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
     3961 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaaggggt ctgacgctca
     4021 gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg tctgctctgc
     4081 aggacgcgtg tccgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca
     4141 ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca
     4201 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     4261 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     4321 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     4381 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     4441 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     4501 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     4561 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
     4621 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     4681 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     4741 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     4801 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     4861 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     4921 gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg
     4981 gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt
     5041 tattcggaca cgcgtcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5101 attttaacaa aatattaacg tttacaattt aaatatttgc ttatacaatc ttcctgtttt
     5161 tggggctttt ctgattatca accggggtac atatgattga catgctagtt ttacgattac
     5221 cgttcatcga ttctcttgtt tgctccagac tctcaggcaa tgacctgata gcctttgtag
     5281 atctctcaaa aatagctacc ctctccggca ttaatttatc agctagaacg gttgaatatc
     5341 atattgatgg tgatttgact gtctccggcc tttctcaccc ttttgaatct ttacctacac
     5401 attactcagg cattgcattt aaaatatatg agggttctaa aaatttttat ccttgcgttg
     5461 aaataaaggc ttctcccgca aaagtattac agggtcataa tgtttttggt acaaccgatt
     5521 tagctttatg ctctgaggct ttattgctta attttgctaa ttctttgcct tgcctgtatg
     5581 atttattgga tgttaatgct actactatta gtagaattga tgccaccttt tcagctcgcg
     5641 ccccaaatga aaatatagct aaacaggtta ttgaccattt gcgaaatgta tctaatggtc
     5701 aaactaaatc tactcgttcg cagaattggg aatcaactgt tatatggaat gaaacttcca
     5761 gacaccgtac tttagttgca tatttaaaac atgttgagct acagcattat attcagcaat
     5821 taagctctaa gccatccgca aaaatgacct cttatcaaaa ggagcaatta aaggtactct
     5881 ctaatcctga cctgttggag tttgcttccg gtctggttcg ctttgaagct cgaattaaaa
     5941 cgcgatattt gaagtctttc gggcttcctc ttaatctttt tgatgcaatc cgctttgctt
     6001 ctgactataa tagtcagggt aaagacctga tttttgattt atggtcattc tcgttttctg
     6061 aactgtttaa agcatttgag ggggattcaa tgaatattta tgacgattcc gcagtattgg
     6121 acgctatcca gtctaaacat tttactatta ccccctctgg caaaacttct tttgcaaaag
     6181 cctctcgcta ttttggtttt tatcgtcgtc tggtaaacga gggttatgat agtgttgctc
     6241 ttactatgcc tcgtaattcc ttttggcgtt atgtatctgc attagttgaa tgtggtattc
     6301 ctaaatctca actgatgaat ctttctacct gtaataatgt tgttccgtta gttcgtttta
     6361 ttaacgtaga tttttcttcc caacgtcctg actggtataa tgagccagtt cttaaaatcg
     6421 cataaggtaa ttcacaatga ttaaagttga aattaaacca tctcaagccc aatttactac
     6481 tcgttctggt gtttctcgtc agggcaagcc ttattcactg aatgagcagc tttgttacgt
     6541 tgatttgggt aatgaatatc cggttcttgt caagattact cttgatgaag gtcagccagc
     6601 ctatgcgcct ggtctgtaca ccgttcatct gtcctctttc aaagttggtc agttcggttc
     6661 ccttatgatt gaccgtctgc gcctcgttcc ggctaagtaa catggagcag gtcgcggatt
     6721 tcgacacaat ttatcaggcg atgatacaaa tctccgttgt actttgtttc gcgcttggta
     6781 taatcgctgg gggtcaaaga tgagtgtttt agtgtattct tttgcctctt tcgttttagg
     6841 ttggtgcctt cgtagtggca ttac
//
