LOCUS       sequence-233904-        4595 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 4595)
  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 4595)
  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-splitD
FEATURES             Location/Qualifiers
     source          1..4595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..1255
                     /codon_start=1
                     /gene="II"
                     /product="pII"
                     /label=M13 gene II
                     /note="replication"
                     /translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
                     TVEYHVDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
                     FGTTDLGLCSEALLLNFANSLPCLYDLLDVNATTISRIDAAFSARAPNENIAKQVIDHL
                     RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
                     KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
                     DLWSFSFSELFKAFEGNSMNIYDDSAVLAAIQSKHFTITPSGKTSFAKASRYFGFYRRL
                     VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
                     DWYNEPVLKIA"
     CDS             1267..1530
                     /codon_start=1
                     /gene="V"
                     /product="pV"
                     /label=M13 gene V
                     /note="replication"
                     /translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQFCYVDLGNEYPVLV
                     KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
     CDS             1532..1633
                     /codon_start=1
                     /gene="VII"
                     /product="pVII"
                     /label=M13 gene VII
                     /note="minor coat protein"
                     /translation="MEQVADFDIIYQAMIQISVVLCFALGIIAGGQR"
     CDS             1630..1728
                     /codon_start=1
                     /gene="IX"
                     /product="pIX"
                     /label=M13 gene IX
                     /note="minor coat protein"
                     /translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
     CDS             1725..1946
                     /codon_start=1
                     /gene="VIII"
                     /product="pVIII"
                     /label=M13 gene VIII
                     /note="major coat protein"
                     /translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFDSLQASATEYI
                     GYAWAMVVVIVGATIGIKLFKKFTSKAS"
     terminator      1953..1992
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(2005..2820)
                     /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(2005..2820)
                     /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     2728..2747
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2812..2831
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(2888..3838)
                     /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      3886..4108
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 cttattatca accggggtac atatgattga catgctagtt ttacggttac cgttcatcga
       61 ttctcttgtt tgctccagac tctctggcaa tgacctgata gcctttgtag atctctcaaa
      121 aatagctacc ctctctggca tgaatttatc agctagaacg gttgaatatc atgttgatgg
      181 tgatttgact gtctccggcc tttcccaccc ttttgaatct ttacctacac attactcagg
      241 cattgcattt aaaatatatg agggttctaa aaatttttat ccttgcgttg aaataaaggc
      301 ttctcccgca aaagtattac aggggcataa tgtttttggt acaaccgatt taggtttatg
      361 ctctgaggct ttattgctta attttgctaa ttctttgcct tgcctgtatg atttattgga
      421 tgttaacgct actactatta gtagaattga tgccgccttt tcagcccgcg ccccaaatga
      481 aaatatagct aaacaggtta ttgaccattt gcgaaatgta tctaatggtc aaactaaatc
      541 tactcgttcg cagaattggg aatcaactgt tacatggaat gaaacttcca gacaccgtac
      601 tttagttgca tatttaaaac atgttgagct acagcaccag attcagcaat taagctctaa
      661 gccatccgca aaaatgacct cttatcaaaa ggaacaatta aaggtactct ctaatcctga
      721 cctgttggaa tttgcttccg gtctggttcg ctttgaggct cgaattaaaa cgagatattt
      781 gaagtctttc gggcttcctc ttaatctttt tgatgcaatc cgctttgctt ctgactataa
      841 tagtcagggt aaagacctga tttttgattt atggtcattc tcgttttctg aactgtttaa
      901 agcatttgag ggaaattcaa tgaatattta tgacgattcc gcagtattgg ccgctatcca
      961 gtctaaacat tttactatta ccccctctgg caaaacttct tttgcaaaag cctctcgcta
     1021 ttttggtttt tatcgtcgtc tggtaaacga gggttatgat agtgttgctc ttactatgcc
     1081 tcgtaattcc ttttggcgtt atgtatctgc attagttgaa tgtggtattc ctaaatctca
     1141 actgatgaat ctttctacct gtaataatgt tgttccgtta gttcgtttta ttaacgtaga
     1201 tttttcttcc caacgtcctg actggtataa tgagccagtt cttaaaatcg cataaggtaa
     1261 ttcacaatga ttaaagttga aattaaacca tctcaagccc aatttactac tcgttctggt
     1321 gtttctcgtc agggcaagcc ttattcactg aatgagcagt tttgttacgt tgatttgggt
     1381 aatgaatatc cggttcttgt caagattact cttgatgaag gtcagccagc ctatgcgccg
     1441 gggctgtaca ctgttcatct gtcctctttc aaagttggtc agttcggttc ccttatgatt
     1501 gaccgtctgc gcctcgttcc ggctaagtaa catggagcag gtcgcggatt tcgacataat
     1561 ttatcaggcg atgatacaaa tctccgttgt actctgtttc gcgcttggta taatagctgg
     1621 gggtcaaaga tgagtgtttt agtgtattct ttcgcctctt tcgttttagg ttggtgcctc
     1681 cgtagtggca ttacgtattt tacccgttta atggaaactt cctcatgaaa aagtctttag
     1741 tcctcaaagc ctctgtagcc gttgctaccc tcgttccgat gctgtctttc gctgctgagg
     1801 gtgacgatcc cgcaaaagcg gcctttgact ccctacaagc ctcagcgacc gaatatatcg
     1861 gttatgcgtg ggcgatggtt gttgtcattg tcggcgcaac tatcggtatc aagctgttta
     1921 agaaattcac ctcgaaagca agttgataaa ctgatacaat taaaggctcc ttttggagcc
     1981 tttttttttg gagtagaaga gctattagaa aaactcatcg agcatcaaat gaaactgcaa
     2041 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     2101 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     2161 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     2221 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     2281 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     2341 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     2401 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     2461 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     2521 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     2581 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     2641 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     2701 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     2761 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     2821 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg aaggccaaat
     2881 aggccgttca gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt
     2941 gcgtgagcca tgagaacgaa ccattgagat catgcttact ttgcatgtca ctcaaaaatt
     3001 ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta
     3061 gtccgttacg taggtaggaa tctgatgtaa tggttgttgg tattttgtca ccattcattt
     3121 ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga
     3181 aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag
     3241 tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt
     3301 agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct
     3361 tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt
     3421 tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa
     3481 gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat
     3541 agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc
     3601 tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt
     3661 tcatcatctg agcgtattgg ttataagtga acgataccgt ccgttctttc cttgtagggt
     3721 tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg
     3781 ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac
     3841 atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat
     3901 aattactagt ccttttcctt tgagttgtgg gtatctgtaa attctgctag acctttgctg
     3961 gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt
     4021 ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa
     4081 gaatagatcc cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa
     4141 ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaaggcttaa
     4201 gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg
     4261 cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg
     4321 aatgggggta aatggcacta caggcgcctt ttatggattc atgcaaggaa actacccata
     4381 atacaagaaa agcccgtcac gggcttctca gggcgtttta tggcgggtct gctatgtggt
     4441 gctatctgac tttttgctgt tcagcagttc ctgccctctg attttccagt ctgaccactt
     4501 cggattatcc cgtgacaggt cattcagact ggctaatgca cccagtaagg cagcggtatc
     4561 atcaacaggc ttacccgtct tactgtcgct cttct
//
