LOCUS       sequence-432463-        6611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Helper plasmid that allows phagemid production upon complementation 
            with a phagemid expressing pIII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6611)
  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 6611)
  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-dg3 hp
FEATURES             Location/Qualifiers
     source          1..6611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             75..1121
                     /codon_start=1
                     /gene="I"
                     /product="pI"
                     /label=M13 gene I
                     /note="phage assembly"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     ITGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSLTYIDLCTVSIKKGN
                     SNEIVKCN"
     CDS             1099..2379
                     /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(2483..2500)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2617..3162)
                     /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(2697..2716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3353..4012)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3927..3946
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4013..4115)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4367..5599
                     /codon_start=1
                     /gene="II"
                     /product="pII"
                     /label=M13 gene II
                     /note="replication"
                     /translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGINLSAR
                     TVEYHIDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
                     FGTTDLALCSEALLLNFANSLPCLYDLLDVNATTISRIDATFSARAPNENIAKQVIDHL
                     RNVSNGQTKSTRSQNWESTVIWNETSRHRTLVAYLKHVELQHYIQQLSSKPSAKMTSYQ
                     KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
                     DLWSFSFSELFKAFEGDSMNIYDDSAVLDAIQSKHFTITPSGKTSFAKASRYFGFYRRL
                     VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
                     DWYNEPVLKIA"
     CDS             5611..5874
                     /codon_start=1
                     /gene="V"
                     /product="pV"
                     /label=M13 gene V
                     /note="replication"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             5876..5977
                     /codon_start=1
                     /gene="VII"
                     /product="pVII"
                     /label=M13 gene VII
                     /note="minor coat protein"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             5974..6072
                     /codon_start=1
                     /gene="IX"
                     /product="pIX"
                     /label=M13 gene IX
                     /note="minor coat protein"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             6069..6290
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     terminator      6294..6341
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             join(6346..6611,1..73)
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
ORIGIN
        1 ctctgtaaag gctgctattt tcatttttga cgttaaacaa aaaatcgttt cttatttgga
       61 ttgggataaa taatatggct gtttattttg taactggcaa attaggctct ggaaagacgc
      121 tcgttagcgt tggtaagatt caggataaaa ttgtagctgg gtgcaaaata gcaactaatc
      181 ttgatttaag gcttcaaaac ctcccgcaag tcgggaggtt cgctaaaacg cctcgcgttc
      241 ttagaatacc ggataagcct tctatatctg atttgcttgc tattgggcgc ggtaatgatt
      301 cctacgatga aaataaaaac ggcttgcttg ttctcgatga gtgcggtact tggtttaata
      361 cccgttcttg gaatgataag gaaagacagc cgattattga ttggtttcta catgctcgta
      421 aattaggatg ggatattatt tttcttgttc aggacttatc tattgttgat aaacaggcgc
      481 gttctgcatt agctgaacat gttgtttatt gtcgtcgtct ggacagaatt actttacctt
      541 ttgtcggtac tttatattct cttattactg gctcgaaaat gcctctgcct aaattacatg
      601 ttggcgttgt taaatatggc gattctcaat taagccctac tgttgagcgt tggctttata
      661 ctggtaagaa tttgtataac gcatatgata ctaaacaggc tttttctagt aattatgatt
      721 ccggtgttta ttcttattta acgccttatt tatcacacgg tcggtatttc aaaccattaa
      781 atttaggtca gaagatgaaa ttaactaaaa tatatttgaa aaagttttct cgcgttcttt
      841 gtcttgcgat tggatttgca tcagcattta catatagtta tataacccaa cctaagccgg
      901 aggttaaaaa ggtagtctct cagacctatg attttgataa attcactatt gactcttctc
      961 agcgtcttaa tctaagctat cgctatgttt tcaaggattc taagggaaaa ttaattaata
     1021 gcgacgattt acagaagcaa ggttattcac tcacatatat tgatttatgt actgtttcca
     1081 ttaaaaaagg taattcaaat gaaattgtta aatgtaatta attttgtttt cttgatgttt
     1141 gtttcatcat cttcttttgc tcaggtaatt gaaatgaata attcgcctct gcgcgatttt
     1201 gtaacttggt attcaaagca atcaggcgaa tccgttattg tttctcccga tgtaaaaggt
     1261 actgttactg tatattcatc tgacgttaaa cctgaaaatc tacgcaattt ctttatttct
     1321 gttttacgtg caaataattt tgatatggta ggttctaacc cttccattat tcagaagtat
     1381 aatccaaaca atcaggatta tattgatgaa ttgccatcat ctgataatca ggaatatgat
     1441 gataattccg ctccttctgg tggtttcttt gttccgcaaa atgataatgt tactcaaact
     1501 tttaaaatta ataacgttcg ggcaaaggat ttaatacgag ttgtcgaatt gtttgtaaag
     1561 tctaatactt ctaaatcctc aaatgtatta tctattgacg gctctaatct attagttgtt
     1621 agtgctccta aagatatttt agataacctt cctcaattcc tttcaactgt tgatttgcca
     1681 actgaccaga tattgattga gggtttgata tttgaggttc agcaaggtga tgctttagat
     1741 ttttcatttg ctgctggctc tcagcgtggc actgttgcag gcggtgttaa tactgaccgc
     1801 ctcacctctg ttttatcttc tgctggtggt tcgttcggta tttttaatgg cgatgtttta
     1861 gggctatcag ttcgcgcatt aaagactaat agccattcaa aaatattgtc tgtgccacgt
     1921 attcttacgc tttcaggtca gaagggttct atctctgttg gccagaatgt cccttttatt
     1981 actggtcgtg tgactggtga atctgccaat gtaaataatc catttcagac gattgagcgt
     2041 caaaatgtag gtatttccat gagcgttttt cctgttgcaa tggctggcgg taatattgtt
     2101 ctggatatta ccagcaaggc cgatagtttg agttcttcta ctcaggcaag tgatgttatt
     2161 actaatcaaa gaagtattgc tacaacggtt aatttgcgtg atggacagac tcttttactc
     2221 ggtggcctca ctgattataa aaacacttct caggattctg gcgtaccgtt cctgtctaaa
     2281 atccctttaa tcggcctcct gtttagctcc cgctctgatt ctaacgagga aagcacgtta
     2341 tacgtgctcg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcctgcag
     2401 agaacatggc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
     2461 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
     2521 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
     2581 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
     2641 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
     2701 ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
     2761 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
     2821 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
     2881 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
     2941 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
     3001 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
     3061 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
     3121 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaag
     3181 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa
     3241 actgtctgct ctgcaggacg cgtgtccgaa tttctgccat tcatccgctt attatcactt
     3301 attcaggcgt agcaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
     3361 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
     3421 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     3481 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
     3541 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
     3601 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
     3661 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
     3721 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
     3781 tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
     3841 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
     3901 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
     3961 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
     4021 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     4081 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg
     4141 cccagggctt cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt
     4201 ccgtcacagg tatttattcg gacacgcgtc ctattggtta aaaaatgagc tgatttaaca
     4261 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaatat ttgcttatac
     4321 aatcttcctg tttttggggc ttttctgatt atcaaccggg gtacatatga ttgacatgct
     4381 agttttacga ttaccgttca tcgattctct tgtttgctcc agactctcag gcaatgacct
     4441 gatagccttt gtagatctct caaaaatagc taccctctcc ggcattaatt tatcagctag
     4501 aacggttgaa tatcatattg atggtgattt gactgtctcc ggcctttctc acccttttga
     4561 atctttacct acacattact caggcattgc atttaaaata tatgagggtt ctaaaaattt
     4621 ttatccttgc gttgaaataa aggcttctcc cgcaaaagta ttacagggtc ataatgtttt
     4681 tggtacaacc gatttagctt tatgctctga ggctttattg cttaattttg ctaattcttt
     4741 gccttgcctg tatgatttat tggatgttaa tgctactact attagtagaa ttgatgccac
     4801 cttttcagct cgcgccccaa atgaaaatat agctaaacag gttattgacc atttgcgaaa
     4861 tgtatctaat ggtcaaacta aatctactcg ttcgcagaat tgggaatcaa ctgttatatg
     4921 gaatgaaact tccagacacc gtactttagt tgcatattta aaacatgttg agctacagca
     4981 ttatattcag caattaagct ctaagccatc cgcaaaaatg acctcttatc aaaaggagca
     5041 attaaaggta ctctctaatc ctgacctgtt ggagtttgct tccggtctgg ttcgctttga
     5101 agctcgaatt aaaacgcgat atttgaagtc tttcgggctt cctcttaatc tttttgatgc
     5161 aatccgcttt gcttctgact ataatagtca gggtaaagac ctgatttttg atttatggtc
     5221 attctcgttt tctgaactgt ttaaagcatt tgagggggat tcaatgaata tttatgacga
     5281 ttccgcagta ttggacgcta tccagtctaa acattttact attaccccct ctggcaaaac
     5341 ttcttttgca aaagcctctc gctattttgg tttttatcgt cgtctggtaa acgagggtta
     5401 tgatagtgtt gctcttacta tgcctcgtaa ttccttttgg cgttatgtat ctgcattagt
     5461 tgaatgtggt attcctaaat ctcaactgat gaatctttct acctgtaata atgttgttcc
     5521 gttagttcgt tttattaacg tagatttttc ttcccaacgt cctgactggt ataatgagcc
     5581 agttcttaaa atcgcataag gtaattcaca atgattaaag ttgaaattaa accatctcaa
     5641 gcccaattta ctactcgttc tggtgtttct cgtcagggca agccttattc actgaatgag
     5701 cagctttgtt acgttgattt gggtaatgaa tatccggttc ttgtcaagat tactcttgat
     5761 gaaggtcagc cagcctatgc gcctggtctg tacaccgttc atctgtcctc tttcaaagtt
     5821 ggtcagttcg gttcccttat gattgaccgt ctgcgcctcg ttccggctaa gtaacatgga
     5881 gcaggtcgcg gatttcgaca caatttatca ggcgatgata caaatctccg ttgtactttg
     5941 tttcgcgctt ggtataatcg ctgggggtca aagatgagtg ttttagtgta ttcttttgcc
     6001 tctttcgttt taggttggtg ccttcgtagt ggcattacgt attttacccg tttaatggaa
     6061 acttcctcat gaaaaagtct ttagtcctca aagcctctgt agccgttgct accctcgttc
     6121 cgatgctgtc tttcgctgct gagggtgacg atcccgcaaa agcggccttt aactccctgc
     6181 aagcctcagc gaccgaatat atcggttatg cgtgggcgat ggttgttgtc attgtcggcg
     6241 caactatcgg tatcaagctg tttaagaaat tcacctcgaa agcaagctga taaaccgata
     6301 caattaaagg ctccttttgg agcctttttt ttggagattt tcaacatgcc agttcttttg
     6361 ggtattccgt tattattgcg tttcctcggt ttccttctgg taactttgtt cggctatctg
     6421 cttacttttc ttaaaaaggg cttcggtaag atagctattg ctatttcatt gtttcttgct
     6481 cttattattg ggcttaactc aattcttgtg ggttatctct ctgatattag cgctcaatta
     6541 ccctctgact ttgttcaggg tgttcagtta attctcccgt ctaatgcgct tccctgtttt
     6601 tatgttattc t
//
