LOCUS       sequence-358519-        4924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phagemid (encodes evolved cI4A5T6T,P 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,P-J23106-geneIII
FEATURES             Location/Qualifiers
     source          1..4924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(164..232,233..1024)
                     /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(382..401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        1048..1082
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1107..2381
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKSHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDLFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     terminator      2396..2467
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2483..2510
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2535..3044)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2730..2751)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2938..2957
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3155..3743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3644..3663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3804..3825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      4780..4851
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4867..4894
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 cgcttcggcg ggcttttttt tgttaactac gtcaggtggc acttttcggg gaaatgtgcg
       61 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      121 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      181 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      241 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      301 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gtttcccaat
      361 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
      421 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      481 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      541 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
      601 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
      661 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
      721 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
      781 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
      841 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcccgcggta tcattgcagc
      901 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
      961 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1021 gtaactgtca gaccaagttt actcatattt acggctagct cagtcctagg tatagtgcta
     1081 gcgaagacaa agatgagaaa tactaggtga aaaaattatt attcgcaatt cctttagttg
     1141 ttcctttcta ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa tcccatacag
     1201 aaaattcatt tactaacgtc tggaaagacg acaaaacttt agatcgttac gctaactatg
     1261 agggctgtct gtggaatgct acaggcgttg tagtttgtac tggtgacgaa actcagtgtt
     1321 acggtacatg ggttcctatt gggcttgcta tccctgaaaa tgagggtggt ggctctgagg
     1381 gtggcggttc tgagggtggc ggttctgagg gtggcggtac taaacctcct gagtacggtg
     1441 atacacctat tccgggctat acttatatca accctctcga cggcacttat ccgcctggta
     1501 ctgagcaaaa ccccgctaat cctaatcctt ctcttgagga gtctcagcct cttaatactt
     1561 tcatgtttca gaataatagg ttccgaaata ggcagggggc attaactgtt tatacgggca
     1621 ctgttactca aggcactgac cccgttaaaa cttattacca gtacactcct gtatcatcaa
     1681 aagccatgta tgacgcttac tggaacggta aattcagaga ctgcgctttc cattctggct
     1741 ttaatgagga tttatttgtt tgtgaatatc aaggccaatc gtctgacctg cctcaacctc
     1801 ctgtcaatgc tggcggcggc tctggtggtg gttctggtgg cggctctgag ggtggtggct
     1861 ctgagggtgg cggttctgag ggtggcggct ctgagggagg cggttccggt ggtggctctg
     1921 gttccggtga ttttgattat gaaaagatgg caaacgctaa taagggggct atgaccgaaa
     1981 atgccgatga aaacgcgcta cagtctgacg ctaaaggcaa acttgattct gtcgctactg
     2041 attacggtgc tgctatcgat ggtttcattg gtgacgtttc cggccttgct aatggtaatg
     2101 gtgctactgg tgattttgct ggctctaatt cccaaatggc tcaagtcggt gacggtgata
     2161 attcaccttt aatgaataat ttccgtcaat atttaccttc cctccctcaa tcggttgaat
     2221 gtcgcccttt tgtctttggc gctggtaaac catatgaatt ttctattgat tgtgacaaaa
     2281 taaacttatt ccgtggtgtc tttgcgtttc ttttatatgt tgccaccttt atgtatgtat
     2341 tttctacgtt tgctaacata ctgcgtaata aggagtctta agaagaccca ggcatcaaat
     2401 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     2461 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatatata
     2521 ctttagattg atttaccccg gttgataatc agaaaagccc caaaaacagg aagattgtat
     2581 aagcaaatat ttaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     2641 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2701 aatagcccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2761 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2821 aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2881 ctaaagggag cccccgattt agagcttgac ggggaaagcg aacgtggcga gaaaggaagg
     2941 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3001 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtaaaagg atctaggtga
     3061 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3121 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3181 tctgctgctt acaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3241 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3301 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3361 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3421 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3481 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3541 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3601 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3661 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3721 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3781 tttgctggcc ttttgctcac atgtaatgtg agttagctca ctcattagaa ttcgctgaga
     3841 gacccgtgct gatcactgac cacaacgtgc gtgaaacact ggcggtttgt gaacgcgctt
     3901 atatcgtcag tcaggggcat ttgatcgccc acggcacgcc tacagaaatc ttacaagacg
     3961 aacacgttaa gcgtgtatac cttggggaag acttcagact ctgatagggt agaagtttgc
     4021 gacgttttag caggagagta cgattctgaa catgagcaca aaaaagaaac cattaacaca
     4081 agagcagctt gaggacgcac gtcgccttaa agcaatttat gaaaaaaaga aaaatgaact
     4141 tggcttatcc caggaattgg tcgcatacga gatggggatg tggcagaacc gcatctgcgc
     4201 tttatttaat ggcatcgcgg cattaaatgc ttataacgcc gcattgcttg caaaaattct
     4261 caaagttagc gttgaagaat ttagcccttc aatcgccaga gaaatctacg agatgtatga
     4321 agcggttagt atgcagccgt cacttagaag tgagtatgag taccctgttt tttctcatgt
     4381 tcaggcaggg atgttctcac ctgagcttag aacctttacc aaaggtgatg cggagagatg
     4441 ggtaagcaca accaaaaaag ccagtgattc tgcattctgg cttgaggttg aaggtaattc
     4501 catgaccgca ccaacaggct ccaagccaag ctttcctgac ggaatgttaa ttctcgttga
     4561 ccctgagcag gctgttgagc caggtgattt ctgcatagcc agacttgggg gtgatgagtt
     4621 taccttcaag aaactgatca gggatagcgg tcaggtgttt ttacaaccac taaacccaca
     4681 gtacccaatg atcccatgca atgagagttg ttccgttgtg gggaaagtta tcgctagtca
     4741 gtggcctgaa gagacgtttg gctgagaaga cccaggcatc aaataaaacg aaaggctcag
     4801 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     4861 cacactggct caccttcggg tgggcctttc tgcgtttata ggtctcctgg cctgcagtaa
     4921 agcc
//
