LOCUS       sequence-437754-        6644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Helper plasmid that allows phagemid production upon complementation 
            with a phagemid expressing pII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6644)
  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 6644)
  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-dg2 hp
FEATURES             Location/Qualifiers
     source          1..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             69..407
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     CDS             409..1455
                     /codon_start=1
                     /gene="I"
                     /product="pI"
                     /label=M13 gene I
                     /note="phage assembly"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     ITGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSQRLNLSYRYVFKDSKGKLINSDDLQKQGYSLTYIDLCTVSIKKGN
                     SNEIVKCN"
     CDS             1433..2713
                     /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(2817..2834)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2951..3496)
                     /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(3031..3050)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3687..4346)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4261..4280
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4347..4449)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4701..4964
                     /codon_start=1
                     /gene="V"
                     /product="pV"
                     /label=M13 gene V
                     /note="replication"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQLCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             4966..5067
                     /codon_start=1
                     /gene="VII"
                     /product="pVII"
                     /label=M13 gene VII
                     /note="minor coat protein"
                     /translation="MEQVADFDTIYQAMIQISVVLCFALGIIAGGQR"
     CDS             5064..5162
                     /codon_start=1
                     /gene="IX"
                     /product="pIX"
                     /label=M13 gene IX
                     /note="minor coat protein"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             5159..5380
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFNSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     terminator      5384..5431
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             join(5436..6644,1..66)
                     /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 tatgttgcca cctttatgta tgtattttct acgtttgcta acatactgcg taataaggag
       61 tcttaatcat gccagttctt ttgggtattc cgttattatt gcgtttcctc ggtttccttc
      121 tggtaacttt gttcggctat ctgcttactt ttcttaaaaa gggcttcggt aagatagcta
      181 ttgctatttc attgtttctt gctcttatta ttgggcttaa ctcaattctt gtgggttatc
      241 tctctgatat tagcgctcaa ttaccctctg actttgttca gggtgttcag ttaattctcc
      301 cgtctaatgc gcttccctgt ttttatgtta ttctctctgt aaaggctgct attttcattt
      361 ttgacgttaa acaaaaaatc gtttcttatt tggattggga taaataatat ggctgtttat
      421 tttgtaactg gcaaattagg ctctggaaag acgctcgtta gcgttggtaa gattcaggat
      481 aaaattgtag ctgggtgcaa aatagcaact aatcttgatt taaggcttca aaacctcccg
      541 caagtcggga ggttcgctaa aacgcctcgc gttcttagaa taccggataa gccttctata
      601 tctgatttgc ttgctattgg gcgcggtaat gattcctacg atgaaaataa aaacggcttg
      661 cttgttctcg atgagtgcgg tacttggttt aatacccgtt cttggaatga taaggaaaga
      721 cagccgatta ttgattggtt tctacatgct cgtaaattag gatgggatat tatttttctt
      781 gttcaggact tatctattgt tgataaacag gcgcgttctg cattagctga acatgttgtt
      841 tattgtcgtc gtctggacag aattacttta ccttttgtcg gtactttata ttctcttatt
      901 actggctcga aaatgcctct gcctaaatta catgttggcg ttgttaaata tggcgattct
      961 caattaagcc ctactgttga gcgttggctt tatactggta agaatttgta taacgcatat
     1021 gatactaaac aggctttttc tagtaattat gattccggtg tttattctta tttaacgcct
     1081 tatttatcac acggtcggta tttcaaacca ttaaatttag gtcagaagat gaaattaact
     1141 aaaatatatt tgaaaaagtt ttctcgcgtt ctttgtcttg cgattggatt tgcatcagca
     1201 tttacatata gttatataac ccaacctaag ccggaggtta aaaaggtagt ctctcagacc
     1261 tatgattttg ataaattcac tattgactct tctcagcgtc ttaatctaag ctatcgctat
     1321 gttttcaagg attctaaggg aaaattaatt aatagcgacg atttacagaa gcaaggttat
     1381 tcactcacat atattgattt atgtactgtt tccattaaaa aaggtaattc aaatgaaatt
     1441 gttaaatgta attaattttg ttttcttgat gtttgtttca tcatcttctt ttgctcaggt
     1501 aattgaaatg aataattcgc ctctgcgcga ttttgtaact tggtattcaa agcaatcagg
     1561 cgaatccgtt attgtttctc ccgatgtaaa aggtactgtt actgtatatt catctgacgt
     1621 taaacctgaa aatctacgca atttctttat ttctgtttta cgtgcaaata attttgatat
     1681 ggtaggttct aacccttcca ttattcagaa gtataatcca aacaatcagg attatattga
     1741 tgaattgcca tcatctgata atcaggaata tgatgataat tccgctcctt ctggtggttt
     1801 ctttgttccg caaaatgata atgttactca aacttttaaa attaataacg ttcgggcaaa
     1861 ggatttaata cgagttgtcg aattgtttgt aaagtctaat acttctaaat cctcaaatgt
     1921 attatctatt gacggctcta atctattagt tgttagtgct cctaaagata ttttagataa
     1981 ccttcctcaa ttcctttcaa ctgttgattt gccaactgac cagatattga ttgagggttt
     2041 gatatttgag gttcagcaag gtgatgcttt agatttttca tttgctgctg gctctcagcg
     2101 tggcactgtt gcaggcggtg ttaatactga ccgcctcacc tctgttttat cttctgctgg
     2161 tggttcgttc ggtattttta atggcgatgt tttagggcta tcagttcgcg cattaaagac
     2221 taatagccat tcaaaaatat tgtctgtgcc acgtattctt acgctttcag gtcagaaggg
     2281 ttctatctct gttggccaga atgtcccttt tattactggt cgtgtgactg gtgaatctgc
     2341 caatgtaaat aatccatttc agacgattga gcgtcaaaat gtaggtattt ccatgagcgt
     2401 ttttcctgtt gcaatggctg gcggtaatat tgttctggat attaccagca aggccgatag
     2461 tttgagttct tctactcagg caagtgatgt tattactaat caaagaagta ttgctacaac
     2521 ggttaatttg cgtgatggac agactctttt actcggtggc ctcactgatt ataaaaacac
     2581 ttctcaggat tctggcgtac cgttcctgtc taaaatccct ttaatcggcc tcctgtttag
     2641 ctcccgctct gattctaacg aggaaagcac gttatacgtg ctcgtcaaag caaccatagt
     2701 acgcgccctg tagcggcgca ttaagcgcct gcagagaaca tggcttcatg tggcaggaga
     2761 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     2821 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     2881 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     2941 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     3001 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     3061 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     3121 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     3181 gactgtatgc acgacccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     3241 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     3301 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     3361 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     3421 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     3481 accaaaacga tctcaagaag atcatcttat taaggggtct gacgctcagt ggaacgaaaa
     3541 ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgctctgcag gacgcgtgtc
     3601 cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcacc aggcgtttaa
     3661 gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt
     3721 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     3781 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     3841 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     3901 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     3961 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     4021 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     4081 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     4141 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     4201 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     4261 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     4321 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     4381 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     4441 tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg
     4501 acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggacacg
     4561 cgtcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4621 tattaacgtt tacaatttaa atatttgctt atacaatctt cctgtttttg gggcttttct
     4681 gattatcaac cggggtacat atgattaaag ttgaaattaa accatctcaa gcccaattta
     4741 ctactcgttc tggtgtttct cgtcagggca agccttattc actgaatgag cagctttgtt
     4801 acgttgattt gggtaatgaa tatccggttc ttgtcaagat tactcttgat gaaggtcagc
     4861 cagcctatgc gcctggtctg tacaccgttc atctgtcctc tttcaaagtt ggtcagttcg
     4921 gttcccttat gattgaccgt ctgcgcctcg ttccggctaa gtaacatgga gcaggtcgcg
     4981 gatttcgaca caatttatca ggcgatgata caaatctccg ttgtactttg tttcgcgctt
     5041 ggtataatcg ctgggggtca aagatgagtg ttttagtgta ttcttttgcc tctttcgttt
     5101 taggttggtg ccttcgtagt ggcattacgt attttacccg tttaatggaa acttcctcat
     5161 gaaaaagtct ttagtcctca aagcctctgt agccgttgct accctcgttc cgatgctgtc
     5221 tttcgctgct gagggtgacg atcccgcaaa agcggccttt aactccctgc aagcctcagc
     5281 gaccgaatat atcggttatg cgtgggcgat ggttgttgtc attgtcggcg caactatcgg
     5341 tatcaagctg tttaagaaat tcacctcgaa agcaagctga taaaccgata caattaaagg
     5401 ctccttttgg agcctttttt ttggagattt tcaacgtgaa aaaattatta ttcgcaattc
     5461 ctttagttgt tcctttctat tctcactccg ctgaaactgt tgaaagttgt ttagcaaaat
     5521 cccatacaga aaattcattt actaacgtct ggaaagacga caaaacttta gatcgttacg
     5581 ctaactatga gggctgtctg tggaatgcta caggcgttgt agtttgtact ggtgacgaaa
     5641 ctcagtgtta cggtacatgg gttcctattg ggcttgctat ccctgaaaat gagggtggtg
     5701 gctctgaggg tggcggttct gagggtggcg gttctgaggg tggcggtact aaacctcctg
     5761 agtacggtga tacacctatt ccgggctata cttatatcaa ccctctcgac ggcacttatc
     5821 cgcctggtac tgagcaaaac cccgctaatc ctaatccttc tcttgaggag tctcagcctc
     5881 ttaatacttt catgtttcag aataataggt tccgaaatag gcagggggca ttaactgttt
     5941 atacgggcac tgttactcaa ggcactgacc ccgttaaaac ttattaccag tacactcctg
     6001 tatcatcaaa agccatgtat gacgcttact ggaacggtaa attcagagac tgcgctttcc
     6061 attctggctt taatgaggat ttatttgttt gtgaatatca aggccaatcg tctgacctgc
     6121 ctcaacctcc tgtcaatgct ggcggcggct ctggtggtgg ttctggtggc ggctctgagg
     6181 gtggtggctc tgagggtggc ggttctgagg gtggcggctc tgagggaggc ggttccggtg
     6241 gtggctctgg ttccggtgat tttgattatg aaaagatggc aaacgctaat aagggggcta
     6301 tgaccgaaaa tgccgatgaa aacgcgctac agtctgacgc taaaggcaaa cttgattctg
     6361 tcgctactga ttacggtgct gctatcgatg gtttcattgg tgacgtttcc ggccttgcta
     6421 atggtaatgg tgctactggt gattttgctg gctctaattc ccaaatggct caagtcggtg
     6481 acggtgataa ttcaccttta atgaataatt tccgtcaata tttaccttcc ctccctcaat
     6541 cggttgaatg tcgccctttt gtctttggcg ctggtaaacc atatgaattt tctattgatt
     6601 gtgacaaaat aaacttattc cgtggtgtct ttgcgtttct ttta
//
