LOCUS       sequence-273119-        4326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phagemid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4326)
  AUTHORS   Brodel AK, Rodrigues R, Jaramillo A, Isalan M
  TITLE     Accelerated evolution of a minimal 63-amino acid dual transcription 
            factor.
  JOURNAL   Sci Adv. 2020 Jun 10;6(24):eaba2728. doi: 10.1126/sciadv.aba2728. 
            eCollection 2020 Jun.
  PUBMED    32577520
REFERENCE   2  (bases 1 to 4326)
  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: "Sci Adv."; 
            date: "2020-06-10"; volume: "6(24):eaba2728. doi"; pages: " 
            10.1126/sciadv.aba2728. eCollection 2020 Jun"
COMMENT     Sequence Label: pLITMUS*-PrpoN-CroAct3-PBBa_J23106-geneIII
FEATURES             Location/Qualifiers
     source          1..4326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        38..142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(143..211,212..1003)
                     /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(361..380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        1027..1061
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             1068..1079
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1086..2360
                     /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      2375..2446
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2462..2489
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2514..3023)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2709..2730)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2917..2936
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3134..3722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3623..3642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3783..3804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gttaactacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
       61 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      121 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      181 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      241 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      301 gatccttgag agttttcgcc ccgaagaacg tttcccaatg atgagcactt ttaaagttct
      361 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
      421 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      481 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      541 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      601 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
      661 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
      721 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      781 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
      841 tggagccggt gagcgtgggt cccgcggtat cattgcagca ctggggccag atggtaagcc
      901 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
      961 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1021 ctcatattta cggctagctc agtcctaggt atagtgctag cgaagacaaa gaggagaaat
     1081 actaggtgaa aaaattatta ttcgcaattc ctttagttgt tcctttctat tctcactccg
     1141 ctgaaactgt tgaaagttgt ttagcaaaat cccatacaga aaattcattt actaacgtct
     1201 ggaaagacga caaaacttta gatcgttacg ctaactatga gggctgtctg tggaatgcta
     1261 caggcgttgt agtttgtact ggtgacgaaa ctcagtgtta cggtacatgg gttcctattg
     1321 ggcttgctat ccctgaaaat gagggtggtg gctctgaggg tggcggttct gagggtggcg
     1381 gttctgaggg tggcggtact aaacctcctg agtacggtga tacacctatt ccgggctata
     1441 cttatatcaa ccctctcgac ggcacttatc cgcctggtac tgagcaaaac cccgctaatc
     1501 ctaatccttc tcttgaggag tctcagcctc ttaatacttt catgtttcag aataataggt
     1561 tccgaaatag gcagggggca ttaactgttt atacgggcac tgttactcaa ggcactgacc
     1621 ccgttaaaac ttattaccag tacactcctg tatcatcaaa agccatgtat gacgcttact
     1681 ggaacggtaa attcagagac tgcgctttcc attctggctt taatgaggat ttatttgttt
     1741 gtgaatatca aggccaatcg tctgacctgc ctcaacctcc tgtcaatgct ggcggcggct
     1801 ctggtggtgg ttctggtggc ggctctgagg gtggtggctc tgagggtggc ggttctgagg
     1861 gtggcggctc tgagggaggc ggttccggtg gtggctctgg ttccggtgat tttgattatg
     1921 aaaagatggc aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac
     1981 agtctgacgc taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg
     2041 gtttcattgg tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg
     2101 gctctaattc ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt
     2161 tccgtcaata tttaccttcc ctccctcaat cggttgaatg tcgccctttt gtctttggcg
     2221 ctggtaaacc atatgaattt tctattgatt gtgacaaaat aaacttattc cgtggtgtct
     2281 ttgcgtttct tttatatgtt gccaccttta tgtatgtatt ttctacgttt gctaacatac
     2341 tgcgtaataa ggagtcttaa gaagacccag gcatcaaata aaacgaaagg ctcagtcgaa
     2401 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac
     2461 tggctcacct tcgggtgggc ctttctgcgt ttatatatac tttagattga tttaccccgg
     2521 ttgataatca gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa
     2581 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2641 aataggccga aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga
     2701 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2761 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt
     2821 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2881 gagcttgacg gggaaagcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2941 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3001 taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa gatccttttt gataatctca
     3061 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3121 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3181 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3241 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3301 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3361 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3421 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3481 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3541 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3601 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3661 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3721 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3781 tgtaatgtga gttagctcac tcattagaat tcgctgagag acccgtgctg atcactgacc
     3841 acaacgtgcg tgaaacactg gcggtttgtg aacgcgctta tatcgtcagt caggggcatt
     3901 tgatcgccca cggcacgcct acagaaatct tacaagacga acacgttaag cgtgtatacc
     3961 ttggggaaga cttcagactc tgatagggta gaagtttgcg acgttttagc aggagagtac
     4021 gattctgaac atggaacaac gcataaccct gaaagattat gcaatgcgct ttgggcaagt
     4081 gaagacagct gcggagctcg gcgtaaacca aagcgcgatc aacaaggcca ttcatgcagg
     4141 ccgaaagatt tttttaacta taaacgctga tggaagcgtt tatgcggaag aggtaaagcc
     4201 cttcccgagt aacaaaaaaa caacagcata gtaagtaatg actaatagcg caaaaaaccc
     4261 cgcttcggcg gggttttttc gcgtctcctg gcctgcagta aagcccgctt cggcgggctt
     4321 tttttt
//
