LOCUS       sequence-243372-        5939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  clone M13 phage by SapI Golden Gate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5939)
  AUTHORS   Thuronyi BW, Koblan LW, Levy JM, Yeh WH, Zheng C, Newby GA, Wilson 
            C, Bhaumik M, Shubina-Oleinik O, Holt JR, Liu DR
  TITLE     Continuous evolution of base editors with expanded target 
            compatibility and improved activity.
  JOURNAL   Nat Biotechnol. 2019 Jul 22. pii: 10.1038/s41587-019-0193-0. doi: 
            10.1038/s41587-019-0193-0.
  PUBMED    31332326
REFERENCE   2  (bases 1 to 5939)
  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 
            Biotechnol. 2019 Jul 22. pii: 10.1038/s41587-019-0193-0. doi: 
            10.1038/s41587-019-0193-0."
COMMENT     Sequence Label: pBT137-splitC
FEATURES             Location/Qualifiers
     source          1..5939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(14..829)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(14..829)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     737..756
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     821..840
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(897..1847)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1895..2117
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2813..3151
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKTAIFIFDVKQKI
                     VSYLDWDK"
     CDS             3153..4199
                     /codon_start=1
                     /gene="I"
                     /product="pI"
                     /label=M13 gene I
                     /note="phage assembly"
                     /translation="MAVYFVTGKLGSGKTLVSVGKIQDKIVAGCKIATNLDLRLQNLPQ
                     VGRFAKTPRVLRIPDKPSISDLLAIGRGNDSYDENKNGLLVLDECGTWFNTRSWNDKER
                     QPIIDWFLHARKLGWDIIFLVQDLSIVDKQARSALAEHVVYCRRLDRITLPFVGTLYSL
                     ITGSKMPLPKLHVGVVKYGDSQLSPTVERWLYTGKNLYNAYDTKQAFSSNYDSGVYSYL
                     TPYLSHGRYFKPLNLGQKMKLTKIYLKKFSRVLCLAIGFASAFTYSYITQPKPEVKKVV
                     SQTYDFDKFTIDSSHRLNLSYRYVFKDSKGKLINSDDLQKQGYSLTYIDLCTVSIKKGN
                     SNEIVKCN"
     CDS             4177..5457
                     /codon_start=1
                     /gene="IV"
                     /product="pIV"
                     /label=M13 gene IV
                     /note="phage assembly"
                     /translation="MKLLNVINFVFLMFVSSSSFAQVIEMNNSPLRDFITWYSKQSGES
                     VIVSPDVKGTVTVYSSDVKSENLRNFFISVLRANNFDMVGSNPSIIQKYNPNNQDYIDE
                     LPSPDNQEYDDNSAPSGGFFVPQNDNVTQTFKINNVRAKDLLRVVELFVKSNTSKSSNV
                     LSIDGSNLLVVSAPKDILDNLSQFLSTVDLPTDQVLIEGLIFEVQQGDALDFSFAAGSQ
                     RGTVAGGVNTDRLTSVLSSAGGSFGIFNGDVLGLSVHALKTNSHSKILSVPRILTLSGQ
                     KGSISIGQNVPFITGRVTGESANVNNPFQTIERQNVGISMSVFPVAMAGGNIVLDITSK
                     ADSLSSSTQASDVITNQRSIATTVNLRDGQTLLLGGLTDYKNTSQDSGVPFLSKIPLIG
                     LLFSSRSDSNEESTLYVLVKATIVRAL"
     rep_origin      5445..5899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5532..5551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5741..5762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cttagaagag ctattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
       61 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
      121 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
      181 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
      241 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
      301 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
      361 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
      421 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
      481 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
      541 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
      601 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
      661 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
      721 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
      781 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg
      841 tattactgtt tatgtaagca gacagtttta ttgttcatga aggccaaata ggccgttcag
      901 atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat
      961 gagaacgaac cattgagatc atgcttactt tgcatgtcac tcaaaaattt tgcctcaaaa
     1021 ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttacgt
     1081 aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg
     1141 ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta
     1201 tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct
     1261 ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta gttattttca
     1321 agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc
     1381 ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa
     1441 gacttaacat gttccagatt atattttatg aattttttta actggaaaag ataaggcaat
     1501 atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata gtttgtccac
     1561 tggaaaatct caaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc
     1621 tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga
     1681 gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg
     1741 gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag
     1801 cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacataca tctcaattgg
     1861 tctaggtgat tttaatcact ataccaattg agatgggcta gtcaatgata attactagtc
     1921 cttttccttt gagttgtggg tatctgtaaa ttctgctaga cctttgctgg aaaacttgta
     1981 aattctgcta gaccctctgt aaattccgct agacctttgt gtgttttttt tgtttatatt
     2041 caagtggtta taatttatag aataaagaaa gaataaaaaa agataaaaag aatagatccc
     2101 agccctgtgt ataactcact actttagtca gttccgcagt attacaaaag gatgtcgcaa
     2161 acgctgtttg ctcctctaca aaacagacct taaaacccta aaggcttaag tagcaccctc
     2221 gcaagctcgg gcaaatcgct gaatattcct tttgtctccg accatcaggc acctgagtcg
     2281 ctgtcttttt cgtgacattc agttcgctgc gctcacggct ctggcagtga atgggggtaa
     2341 atggcactac aggcgccttt tatggattca tgcaaggaaa ctacccataa tacaagaaaa
     2401 gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg ctatctgact
     2461 ttttgctgtt cagcagttcc tgccctctga ttttccagtc tgaccacttc ggattatccc
     2521 gtgacaggtc attcagactg gctaatgcac ccagtaaggc agcggtatca tcaacaggct
     2581 tacccgtctt actgtcgctc ttctggctct agaaggagat attcaacatg ctccctcaat
     2641 cggttgaatg tcgccctttt gtctttggcg ctggtaaacc atatgaattt tctattgatt
     2701 gtgacaaaat aaacttattc cgtggtgttt ttgcgtttct tttatatgtt gccaccttta
     2761 tgtatgtatc ttctacgttt gctaacatac tgcgtaataa ggagtcttaa tcatgccagt
     2821 tcttttgggt attccgttat tattgcgttt cctcggtttc cttctggtaa ctttgttcgg
     2881 ctatctgctt acttttctca aaaagggctt cggtaagata gctattgcta tttcattgtt
     2941 tcttgctctt attattgggc ttaactcaat tcttgtgggt tatctctctg atattagtgc
     3001 tcaattaccc tctgactttg ttcagggtgt tcagttaatt ctcccgtcta atgcgcttcc
     3061 ctgtttttat gttattcttt ctgtaaagac tgctattttc atttttgacg ttaaacaaaa
     3121 aatcgtttct tatttggatt gggacaaata atatggctgt ttattttgta actggcaaat
     3181 taggctctgg aaagacgctc gttagcgttg gtaagattca ggataaaatt gtagctgggt
     3241 gcaaaatagc aactaatctt gatttaaggc ttcaaaacct cccgcaagtc gggaggttcg
     3301 ctaaaacgcc tcgcgttctt agaataccgg ataagccttc tatatctgat ttgcttgcta
     3361 ttgggcgcgg taatgattcc tacgatgaaa ataaaaacgg attgcttgtt ctcgatgagt
     3421 gcggtacttg gtttaatacc cgttcttgga atgataagga aagacagccg attattgatt
     3481 ggtttctaca tgctcgtaaa ttaggatggg atattatttt tcttgttcag gacttatcta
     3541 ttgttgataa acaggcgcgt tctgcattag ctgaacatgt tgtttattgt cgtcgtctgg
     3601 acagaattac tttacctttt gtcggtactt tatattctct tattactggc tcgaaaatgc
     3661 ctctgcctaa attacatgtt ggcgttgtta aatatggcga ttctcaatta agccctactg
     3721 ttgagcgttg gctttatact ggtaagaatt tgtataacgc atacgatact aaacaggctt
     3781 tttctagtaa ttatgattcc ggtgtttatt cttatttaac gccttattta tcacacggtc
     3841 ggtatttcaa accattaaat ttaggtcaga agatgaaatt aactaaaata tatttgaaaa
     3901 agttttctcg cgttctttgt cttgcgattg gatttgcatc agcatttaca tatagttata
     3961 taacccaacc taagccggag gttaaaaagg tagtctctca gacctatgat tttgataaat
     4021 tcactattga ctcttctcat cgtcttaatc taagctatcg ctatgttttc aaggattcta
     4081 agggaaaatt aattaatagc gacgatttac agaagcaagg ttattcactt acatatattg
     4141 atttatgtac tgtttccatt aaaaaaggta attcaaatga aattgttaaa tgtaattaat
     4201 tttgttttct tgatgtttgt ttcatcatct tcttttgctc aggtaattga aatgaataat
     4261 tcgcctctgc gcgattttat aacttggtat tcaaagcaat caggcgaatc cgttattgtt
     4321 tctcccgatg taaaaggtac tgttactgta tattcatctg acgttaaatc tgaaaatcta
     4381 cgcaatttct ttatttctgt tttacgtgca aataattttg atatggtagg ttctaaccct
     4441 tccattattc agaagtataa tccaaacaat caggattata ttgatgaatt gccatcacct
     4501 gataatcagg aatatgatga taattccgct ccttctggtg gtttctttgt cccgcaaaat
     4561 gataatgtta ctcaaacttt taaaattaat aacgttcggg caaaggattt actacgagtt
     4621 gtcgaattgt ttgtaaagtc taatacttct aaatcctcaa atgtattatc tattgacgga
     4681 tctaatctat tagttgttag tgctcctaaa gatattttag ataacctttc tcaattcctt
     4741 tcaactgttg atttgccaac tgaccaggta ttgattgagg gtttgatatt tgaggttcag
     4801 caaggtgatg ctttagattt ttcatttgct gctggctctc agcgtggcac tgttgcaggc
     4861 ggagttaata ctgaccgcct cacctctgtt ttatcttctg ctggtggttc gttcggtatt
     4921 tttaatggcg atgttttagg gctatcagtt cacgcattaa agactaatag ccattcaaaa
     4981 atattgtctg tgccacgtat tcttacgctt tcaggtcaga agggttctat ctctattggc
     5041 cagaatgtcc cttttattac tggtcgtgta actggtgaat ctgccaatgt aaataatcca
     5101 tttcagacga ttgagcgtca aaatgtaggt atttccatga gcgtttttcc tgttgcaatg
     5161 gctggcggta atattgttct ggatattacc agcaaggccg atagtttgag ttcttctact
     5221 caggcaagtg atgttattac taaccaaaga agtattgcta caacggttaa tttgcgtgat
     5281 ggacagactc ttctactcgg tggcctcact gattataaaa acacttctca ggattctggc
     5341 gtaccgttcc tgtctaaaat ccctttaatc ggcctcctgt ttagctcccg ctctgactct
     5401 aacgaggaga gcacgttata cgtgctcgtc aaagcaacca tagtacgcgc cctgtagcgg
     5461 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5521 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5581 ccgtcaagcg ctaaatcggg ggccccttta gggttccgat ttagtgcttt acggcacctc
     5641 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5701 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5761 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     5821 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5881 atattaacgt ttacaattta aatatttgct tatacaatct tcctgttttt ggggctttt
//
