LOCUS       sequence-432465-        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             591..929
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     CDS             931..2211
                     /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(2315..2332)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2449..2994)
                     /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(2529..2548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3185..3844)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3759..3778
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3845..3947)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4199..5431
                     /codon_start=1
                     /gene="II"
                     /product="pII"
                     /label=M13 gene II
                     /note="replication"
                     /translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGINLSAR
                     TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
                     FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
                     RNVSNGQTKSTRSQNWESTVIWNETSRHRTLVAYLKHVELQHYIQQLSSKPSAKMTSYQ
                     KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
                     DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
                     VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
                     DWYNEPVLKIA"
     CDS             5443..5706
                     /codon_start=1
                     /gene="V"
                     /product="pV"
                     /label=M13 gene V
                     /note="replication"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             5708..5809
                     /codon_start=1
                     /gene="VII"
                     /product="pVII"
                     /label=M13 gene VII
                     /note="minor coat protein"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             5806..5904
                     /codon_start=1
                     /gene="IX"
                     /product="pIX"
                     /label=M13 gene IX
                     /note="minor coat protein"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             5901..6122
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     terminator      6126..6173
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             join(6178..6864,1..588)
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKSHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDLFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
ORIGIN
        1 caacctcctg tcaatgctgg cggcggctct ggtggtggtt ctggtggcgg ctctgagggt
       61 ggtggctctg agggtggcgg ttctgagggt ggcggctctg agggaggcgg ttccggtggt
      121 ggctctggtt ccggtgattt tgattatgaa aagatggcaa acgctaataa gggggctatg
      181 accgaaaatg ccgatgaaaa cgcgctacag tctgacgcta aaggcaaact tgattctgtc
      241 gctactgatt acggtgctgc tatcgatggt ttcattggtg acgtttccgg ccttgctaat
      301 ggtaatggtg ctactggtga ttttgctggc tctaattccc aaatggctca agtcggtgac
      361 ggtgataatt cacctttaat gaataatttc cgtcaatatt taccttccct ccctcaatcg
      421 gttgaatgtc gcccttttgt ctttggcgct ggtaaaccat atgaattttc tattgattgt
      481 gacaaaataa acttattccg tggtgtcttt gcgtttcttt tatatgttgc cacctttatg
      541 tatgtatttt ctacgtttgc taacatactg cgtaataagg agtcttaatc atgccagttc
      601 ttttgggtat tccgttatta ttgcgtttcc tcggtttcct tctggtaact ttgttcggct
      661 atctgcttac ttttcttaaa aagggcttcg gtaagatagc tattgctatt tcattgtttc
      721 ttgctcttat tattgggctt aactcaattc ttgtgggtta tctctctgat attagcgctc
      781 aattaccctc tgactttgtt cagggtgttc agttaattct cccgtctaat gcgcttccct
      841 gtttttatgt tattctctct gtaaaggctg ctattttcat ttttgacgtt aaacaaaaaa
      901 tcgtttctta tttggattgg gataaataat atgaaattgt taaatgtaat taattttgtt
      961 ttcttgatgt ttgtttcatc atcttctttt gctcaggtaa ttgaaatgaa taattcgcct
     1021 ctgcgcgatt ttgtaacttg gtattcaaag caatcaggcg aatccgttat tgtttctccc
     1081 gatgtaaaag gtactgttac tgtatattca tctgacgtta aacctgaaaa tctacgcaat
     1141 ttctttattt ctgttttacg tgcaaataat tttgatatgg taggttctaa cccttccatt
     1201 attcagaagt ataatccaaa caatcaggat tatattgatg aattgccatc atctgataat
     1261 caggaatatg atgataattc cgctccttct ggtggtttct ttgttccgca aaatgataat
     1321 gttactcaaa cttttaaaat taataacgtt cgggcaaagg atttaatacg agttgtcgaa
     1381 ttgtttgtaa agtctaatac ttctaaatcc tcaaatgtat tatctattga cggctctaat
     1441 ctattagttg ttagtgctcc taaagatatt ttagataacc ttcctcaatt cctttcaact
     1501 gttgatttgc caactgacca gatattgatt gagggtttga tatttgaggt tcagcaaggt
     1561 gatgctttag atttttcatt tgctgctggc tctcagcgtg gcactgttgc aggcggtgtt
     1621 aatactgacc gcctcacctc tgttttatct tctgctggtg gttcgttcgg tatttttaat
     1681 ggcgatgttt tagggctatc agttcgcgca ttaaagacta atagccattc aaaaatattg
     1741 tctgtgccac gtattcttac gctttcaggt cagaagggtt ctatctctgt tggccagaat
     1801 gtccctttta ttactggtcg tgtgactggt gaatctgcca atgtaaataa tccatttcag
     1861 acgattgagc gtcaaaatgt aggtatttcc atgagcgttt ttcctgttgc aatggctggc
     1921 ggtaatattg ttctggatat taccagcaag gccgatagtt tgagttcttc tactcaggca
     1981 agtgatgtta ttactaatca aagaagtatt gctacaacgg ttaatttgcg tgatggacag
     2041 actcttttac tcggtggcct cactgattat aaaaacactt ctcaggattc tggcgtaccg
     2101 ttcctgtcta aaatcccttt aatcggcctc ctgtttagct cccgctctga ttctaacgag
     2161 gaaagcacgt tatacgtgct cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     2221 aagcgcctgc agagaacatg gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
     2281 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
     2341 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     2401 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     2461 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     2521 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     2581 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     2641 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     2701 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     2761 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     2821 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     2881 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     2941 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     3001 catcttatta aggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3061 atgaacaata aaactgtctg ctctgcagga cgcgtgtccg aatttctgcc attcatccgc
     3121 ttattatcac ttattcaggc gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa
     3181 aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     3241 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     3301 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     3361 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     3421 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     3481 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     3541 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     3601 gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa
     3661 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     3721 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     3781 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     3841 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     3901 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     3961 gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg
     4021 cgaagtgatc ttccgtcaca ggtatttatt cggacacgcg tcctattggt taaaaaatga
     4081 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caatttaaat
     4141 atttgcttat acaatcttcc tgtttttggg gcttttctga ttatcaaccg gggtacatat
     4201 gattgacatg ctagttttac gattaccgtt catcgattct cttgtttgct ccagactctc
     4261 aggcaatgac ctgatagcct ttgtagatct ctcaaaaata gctaccctct ccggcattaa
     4321 tttatcagct agaacggttg aatatcatat tgatggtgat ttgactgtct ccggcctttc
     4381 tcaccctttt gaatctttac ctacacatta ctcaggcatt gcatttaaaa tatatgaggg
     4441 ttctaaaaat ttttatcctt gcgttgaaat aaaggcttct cccgcaaaag tattacaggg
     4501 tcataatgtt tttggtacaa ccgatttagc tttatgctct gaggctttat tgcttaattt
     4561 tgctaattct ttgccttgcc tgtatgattt attggatgtt aatgctacta ctattagtag
     4621 aattgatgcc accttttcag ctcgcgcccc aaatgaaaat atagctaaac aggttattga
     4681 ccatttgcga aatgtatcta atggtcaaac taaatctact cgttcgcaga attgggaatc
     4741 aactgttata tggaatgaaa cttccagaca ccgtacttta gttgcatatt taaaacatgt
     4801 tgagctacag cattatattc agcaattaag ctctaagcca tccgcaaaaa tgacctctta
     4861 tcaaaaggag caattaaagg tactctctaa tcctgacctg ttggagtttg cttccggtct
     4921 ggttcgcttt gaagctcgaa ttaaaacgcg atatttgaag tctttcgggc ttcctcttaa
     4981 tctttttgat gcaatccgct ttgcttctga ctataatagt cagggtaaag acctgatttt
     5041 tgatttatgg tcattctcgt tttctgaact gtttaaagca tttgaggggg attcaatgaa
     5101 tatttatgac gattccgcag tattggacgc tatccagtct aaacatttta ctattacccc
     5161 ctctggcaaa acttcttttg caaaagcctc tcgctatttt ggtttttatc gtcgtctggt
     5221 aaacgagggt tatgatagtg ttgctcttac tatgcctcgt aattcctttt ggcgttatgt
     5281 atctgcatta gttgaatgtg gtattcctaa atctcaactg atgaatcttt ctacctgtaa
     5341 taatgttgtt ccgttagttc gttttattaa cgtagatttt tcttcccaac gtcctgactg
     5401 gtataatgag ccagttctta aaatcgcata aggtaattca caatgattaa agttgaaatt
     5461 aaaccatctc aagcccaatt tactactcgt tctggtgttt ctcgtcaggg caagccttat
     5521 tcactgaatg agcagctttg ttacgttgat ttgggtaatg aatatccggt tcttgtcaag
     5581 attactcttg atgaaggtca gccagcctat gcgcctggtc tgtacaccgt tcatctgtcc
     5641 tctttcaaag ttggtcagtt cggttccctt atgattgacc gtctgcgcct cgttccggct
     5701 aagtaacatg gagcaggtcg cggatttcga cacaatttat caggcgatga tacaaatctc
     5761 cgttgtactt tgtttcgcgc ttggtataat cgctgggggt caaagatgag tgttttagtg
     5821 tattcttttg cctctttcgt tttaggttgg tgccttcgta gtggcattac gtattttacc
     5881 cgtttaatgg aaacttcctc atgaaaaagt ctttagtcct caaagcctct gtagccgttg
     5941 ctaccctcgt tccgatgctg tctttcgctg ctgagggtga cgatcccgca aaagcggcct
     6001 ttaactccct gcaagcctca gcgaccgaat atatcggtta tgcgtgggcg atggttgttg
     6061 tcattgtcgg cgcaactatc ggtatcaagc tgtttaagaa attcacctcg aaagcaagct
     6121 gataaaccga tacaattaaa ggctcctttt ggagcctttt ttttggagat tttcaacgtg
     6181 aaaaaattat tattcgcaat tcctttagtt gttcctttct attctcactc cgctgaaact
     6241 gttgaaagtt gtttagcaaa atcccataca gaaaattcat ttactaacgt ctggaaagac
     6301 gacaaaactt tagatcgtta cgctaactat gagggctgtc tgtggaatgc tacaggcgtt
     6361 gtagtttgta ctggtgacga aactcagtgt tacggtacat gggttcctat tgggcttgct
     6421 atccctgaaa atgagggtgg tggctctgag ggtggcggtt ctgagggtgg cggttctgag
     6481 ggtggcggta ctaaacctcc tgagtacggt gatacaccta ttccgggcta tacttatatc
     6541 aaccctctcg acggcactta tccgcctggt actgagcaaa accccgctaa tcctaatcct
     6601 tctcttgagg agtctcagcc tcttaatact ttcatgtttc agaataatag gttccgaaat
     6661 aggcaggggg cattaactgt ttatacgggc actgttactc aaggcactga ccccgttaaa
     6721 acttattacc agtacactcc tgtatcatca aaagccatgt atgacgctta ctggaacggt
     6781 aaattcagag actgcgcttt ccattctggc tttaatgagg atttatttgt ttgtgaatat
     6841 caaggccaat cgtctgacct gcct
//
