LOCUS       sequence-369552-        4924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phagemid (encodes cI4A5T6T and M13 gene III).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4924)
  AUTHORS   Brodel AK, Jaramillo A, Isalan M
  TITLE     Engineering orthogonal dual transcription factors for multi-input 
            synthetic promoters.
  JOURNAL   Nat Commun. 2016 Dec 16;7:13858. doi: 10.1038/ncomms13858.
  PUBMED    27982027
REFERENCE   2  (bases 1 to 4924)
  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: "Nat 
            Commun."; date: "2016-12-16"; volume: "7:13858. doi"; pages: " 
            10.1038/ncomms13858"
COMMENT     Sequence Label: pLITMUS-rpoN-cI4A5T6T-J23106-geneIII
FEATURES             Location/Qualifiers
     source          1..4924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      993..1064
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1080..1107
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1132..1641)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1327..1348)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1535..1554
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1752..2340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2241..2260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2401..2422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      3377..3448
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3464..3491
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3580..3684
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3685..3753,3754..4545)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3903..3922)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        4569..4603
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             join(4628..4924,1..978)
                     /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 tctgagggtg gcggtactaa acctcctgag tacggtgata cacctattcc gggctatact
       61 tatatcaacc ctctcgacgg cacttatccg cctggtactg agcaaaaccc cgctaatcct
      121 aatccttctc ttgaggagtc tcagcctctt aatactttca tgtttcagaa taataggttc
      181 cgaaataggc agggggcatt aactgtttat acgggcactg ttactcaagg cactgacccc
      241 gttaaaactt attaccagta cactcctgta tcatcaaaag ccatgtatga cgcttactgg
      301 aacggtaaat tcagagactg cgctttccat tctggcttta atgaggattt atttgtttgt
      361 gaatatcaag gccaatcgtc tgacctgcct caacctcctg tcaatgctgg cggcggctct
      421 ggtggtggtt ctggtggcgg ctctgagggt ggtggctctg agggtggcgg ttctgagggt
      481 ggcggctctg agggaggcgg ttccggtggt ggctctggtt ccggtgattt tgattatgaa
      541 aagatggcaa acgctaataa gggggctatg accgaaaatg ccgatgaaaa cgcgctacag
      601 tctgacgcta aaggcaaact tgattctgtc gctactgatt acggtgctgc tatcgatggt
      661 ttcattggtg acgtttccgg ccttgctaat ggtaatggtg ctactggtga ttttgctggc
      721 tctaattccc aaatggctca agtcggtgac ggtgataatt cacctttaat gaataatttc
      781 cgtcaatatt taccttccct ccctcaatcg gttgaatgtc gcccttttgt ctttggcgct
      841 ggtaaaccat atgaattttc tattgattgt gacaaaataa acttattccg tggtgtcttt
      901 gcgtttcttt tatatgttgc cacctttatg tatgtatttt ctacgtttgc taacatactg
      961 cgtaataagg agtcttaaga agacccaggc atcaaataaa acgaaaggct cagtcgaaag
     1021 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     1081 gctcaccttc gggtgggcct ttctgcgttt atatatactt tagattgatt taccccggtt
     1141 gataatcaga aaagccccaa aaacaggaag attgtataag caaatattta aattgtaaac
     1201 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     1261 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat agggttgagt
     1321 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     1381 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt
     1441 ttggggtcga ggtgccgtaa agcactaaat tggaacccta aagggagccc ccgatttaga
     1501 gcttgacggg gaaagcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1561 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1621 atgcgccgct acagggcgcg taaaaggatc taggtgaaga tcctttttga taatctcatg
     1681 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1741 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttaca aacaaaaaaa
     1801 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1861 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1921 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1981 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2041 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2101 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2161 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2221 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2281 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2341 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2401 taatgtgagt tagctcactc attagaattc gctgagagac ccgtgctgat cactgaccac
     2461 aacgtgcgtg aaacactggc ggtttgtgaa cgcgcttata tcgtcagtca ggggcatttg
     2521 atcgcccacg gcacgcctac agaaatctta caagacgaac acgttaagcg tgtatacctt
     2581 ggggaagact tcagactctg atagggtaga agtttgcgac gttttagcag gagagtacga
     2641 ttctgaacat gagcacaaaa aagaaaccat taacacaaga gcagcttgag gacgcacgtc
     2701 gccttaaagc aatttatgaa aaaaagaaaa atgaacttgg cttatcccag gaatctgtcg
     2761 cagacaagat ggggatgtgg cagaaccgca tctgcgcttt atttaatggc atcgcggcat
     2821 taaatgctta taacgccgca ttgcttgcaa aaattctcaa agttagcgtt gaagaattta
     2881 gcccttcaat cgccagagaa atctacgaga tgtatgaagc ggttagtatg cagccgtcac
     2941 ttagaagtga gtatgagtac cctgtttttt ctcatgttca ggcagggatg ttctcacctg
     3001 agcttagaac ctttaccaaa ggtgatgcgg agagatgggt aagcacaacc aaaaaagcca
     3061 gtgattctgc attctggctt gaggttgaag gtaattccat gaccgcacca acaggctcca
     3121 agccaagctt tcctgacgga atgttaattc tcgttgaccc tgagcaggct gttgagccag
     3181 gtgatttctg catagccaga cttgggggtg atgagtttac cttcaagaaa ctgatcaggg
     3241 atagcggtca ggtgttttta caaccactaa acccacagta cccaatgatc ccatgcaatg
     3301 agagttgttc cgttgtgggg aaagttatcg ctagtcagtg gcctgaagag acgtttggct
     3361 gagaagaccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     3421 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     3481 gcctttctgc gtttataggt ctcctggcct gcagtaaagc ccgcttcggc gggctttttt
     3541 ttgttaacta cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3601 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3661 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3721 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3781 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3841 aagatccttg agagttttcg ccccgaagaa cgtttcccaa tgatgagcac ttttaaagtt
     3901 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     3961 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4021 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4081 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4141 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4201 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4261 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4321 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4381 tctggagccg gtgagcgtgg gtcccgcggt atcattgcag cactggggcc agatggtaag
     4441 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4501 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4561 tactcatatt tacggctagc tcagtcctag gtatagtgct agcgaagaca aagatgagaa
     4621 atactaggtg aaaaaattat tattcgcaat tcctttagtt gttcctttct attctcactc
     4681 cgctgaaact gttgaaagtt gtttagcaaa atcccataca gaaaattcat ttactaacgt
     4741 ctggaaagac gacaaaactt tagatcgtta cgctaactat gagggctgtc tgtggaatgc
     4801 tacaggcgtt gtagtttgta ctggtgacga aactcagtgt tacggtacat gggttcctat
     4861 tgggcttgct atccctgaaa atgagggtgg tggctctgag ggtggcggtt ctgagggtgg
     4921 cggt
//
