LOCUS       sequence-478141-        6405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KRAS G12D PANCS-binder positive AP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6405)
  AUTHORS   Styles MJ, Pixley JA, Wei T, Basile C, Lu SS, Dickinson BC
  TITLE     High-throughput protein binder discovery by rapid in vivo selection.
  JOURNAL   bioRxiv [Preprint]. 2025 Jan 17:2025.01.06.631531. doi: 
            10.1101/2025.01.06.631531.
  PUBMED    39829796
REFERENCE   2  (bases 1 to 6405)
  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: "bioRxiv 
            [Preprint]. 2025 Jan 17:2025.01.06.631531. doi: 
            10.1101/2025.01.06.631531."
COMMENT     Sequence Label: 73-21 KRAS PANCS-binder posAP
FEATURES             Location/Qualifiers
     source          1..6405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     160..178
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      230..775
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     676..695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     892..909
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             join(2594..2626,2627..2629,2630..3157)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             2594..3157
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             complement(join(5532..6323,6324..6392))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6155..6174
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6393..6405,1..92))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
       61 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
      121 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
      181 ttaggccaaa taggccgtta gcgtcagacc ccttaataag atgatcttct tgagatcgtt
      241 ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt
      301 cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga ggagcgcagt
      361 caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct
      421 ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac
      481 tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata
      541 cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa
      601 acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac
      661 gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacca
      721 ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg
      781 ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc caggaaatct
      841 ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca
      901 gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac cggtgcagcc
      961 ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca acatagtaag
     1021 ccagtataca ctccgctagc gctaagagga catccggtga aataaaacga aaggctcagt
     1081 cgaaagactg ggcctttcgt tttgctgagg agacttaggg accctactaa taccggtcac
     1141 tatagggaga aagacctgca ggtgcagtaa ggaggaaaaa aaaatgaaaa aattattatt
     1201 cgcaattcct ttagttgttc ctttctattc tcactccgct gaaactgttg aaagttgttt
     1261 agcaaaaccc catacagaaa attcatttac taacgtctgg aaagacgaca aaactttaga
     1321 tcgttacgct aactatgagg gctgtctgtg gaatgctaca ggcgttgtag tttgtactgg
     1381 tgacgaaact cagtgttacg gtacatgggt tcctattggg cttgctatcc ctgaaaatga
     1441 gggtggtggc tctgagggtg gcggttctga gggtggcggt tctgagggtg gcggtactaa
     1501 acctcctgag tacggtgata cacctattcc gggctatact tatatcaacc ctctcgacgg
     1561 cacttatccg cctggtactg agcaaaaccc cgctaatcct aatccttctc ttgaggagtc
     1621 tcagcctctt aatactttca tgtttcagaa taataggttc cgaaataggc agggggcatt
     1681 aactgtttat acgggcactg ttactcaagg cactgacccc gttaaaactt attaccagta
     1741 cactcctgta tcatcaaaag ccatgtatga cgcttactgg aacggtaaat tcagagactg
     1801 cgctttccat tctggcttta atgaggatcc attcgtttgt gaatatcaag gccaatcgtc
     1861 tgacctgcct caacctcctg tcaatgctgg cggcggctct ggtggtggtt ctggtggcgg
     1921 ctctgagggt ggtggctctg agggtggcgg ttctgagggt ggcggctctg agggaggcgg
     1981 ttccggtggt ggctctggtt ccggtgattt tgattatgaa aagatggcaa acgctaataa
     2041 gggggctatg accgaaaatg ccgatgaaaa cgcgctacag tctgacgcta aaggcaaact
     2101 tgattctgtc gctactgatt acggtgctgc tatcgatggt ttcattggtg acgtttccgg
     2161 ccttgctaat ggtaatggtg ctactggtga ttttgctggc tctaattccc aaatggctca
     2221 agtcggtgac ggtgataatt cacctttaat gaataatttc cgtcaatatt taccttccct
     2281 tccacaatcc gttgagtgcc gtccgttcgt gtttggagct ggcaaaccct atgagttcag
     2341 cattgattgt gataagatca atctgtttcg tggggtattt gccttcttat tatatgtcgc
     2401 aacattcatg tatgtgttta gcacattcgc gaacattctt cgcaataaag aatcctaata
     2461 ctagagtatc tgttagtttt tttctactag agtatctgtt agtttttttc catcgtggcc
     2521 ggagtactta agaagataac taaacaacta acggacaatt ctacctagga tccctcgaaa
     2581 ggaggaaaaa aaaatgactg aatataaact tgtggtagtt ggagctgatg gcgtaggcaa
     2641 gagtgccttg acgatacagc taattcagaa tcattttgtg gacgaatatg atccaacaat
     2701 agaggattcc tacaggaagc aagtagtaat tgatggagaa acctgtctct tggatattct
     2761 cgacacagca ggtcaagagg agtacagtgc aatgagggac cagtacatga ggactgggga
     2821 gggctttctt tgtgtatttg ccataaataa tactaaatca tttgaagata ttcaccatta
     2881 tagagaacaa attaaaagag ttaaggactc tgaagatgta cctatggtcc tagtaggaaa
     2941 taaatgtgat ttgccttcta gaacagtaga cacaaaacag gctcaggact tagcaagaag
     3001 ttatggaatt ccttttattg aaacatcagc aaagacaaga cagggtgttg atgatgcctt
     3061 ctatacatta gttcgagaaa ttcgaaaaca taaagaaaag atgagcaaag atggtaaaaa
     3121 gaagaaaaag aagtcaaaga caaagtgtgt aattatgacg tcgggtggaa gcggtggttc
     3181 agggtcctca ggttcgggtg gaactagtgg ctctggtggg tcggggacag gcacagggac
     3241 cggaagtagc ggatcgggtg gcacaagtgg gtctggcggc tctggatcag gcaccggcag
     3301 tgggtcttcg gggagtggag gtacctcagg ctcaggaaaa gcatttatgc aagttgtcga
     3361 ggctgacatg ctctctaagg gtctactcgg tggcgaggcg tggtcttcgt ggcataagga
     3421 agactctatt catgtaggag tacgctgcat cgagatgctc attgagtcaa ccggaatggt
     3481 tagcttacac cgccaaaatg ctggcgtagt aggtcaagac tctgagacta tcgaactcgc
     3541 acctgaatac gctgaggcta tcgcaacccg tgcaggtgcg ctggctggca tctctccgat
     3601 gttccaacct tgcgtagttc ctcctaagcc gtggactggc attactggtg gtggctattg
     3661 ggctaacggt cgtcgtcctc tggcgctggt gcgtactcac agtaagaaag cactgatgcg
     3721 ctacgaagac gtttacatgc ctgaggtgta caaagcgatt aacattgcgc aaaacaccgc
     3781 atggaaaatc aacaagaaag tcctagcggt cgccaacgta atcaccaagt ggaagcattg
     3841 tccggtcgag gacatccctg cgattgagcg tgaagaactc ccgatgaaac cggaagacat
     3901 cgacatgaat cctgaggctc tcaccgcgtg gaaacgtgct gccgctgctg tgtaccgcaa
     3961 ggacaaggct cgcaagtctc gccgtatcag ccttgagttc atgcttgagc aagccaataa
     4021 gtttgctaac cataaggcca tctggttccc ttacaacatg gactggcgcg gtcgtgttta
     4081 cgctgtgtca atgttcaacc cgcaaggtaa cgatatgacc aaaggactgc ttacgctggc
     4141 gaaaggtaaa ccaatcggta aggaaggtta ctactggctg aaaatccacg gtgcaaactg
     4201 tgcgggtgtc gataaggttc cgttccctga gcgcatcaag ttcattgagg aaaaccacga
     4261 gaacatcatg gcttgcgcta agtctccact ggagaacact tggtgggctg agcaagattc
     4321 tccgttctgc ttccttgcgt tctgctttga gtacgctggg gtacagcacc acggcctgag
     4381 ctataactgc tcccttccgc tggcgtttga cgggtcttgc tctggcatcc agcacttctc
     4441 cgcgatgctc cgagatgagg taggtggtcg cgcggttaac ttgcttccta gtgaaaccgt
     4501 tcaggacatc tacgggattg ttgctaagaa agtcaacgag attctacaag cagacgcaat
     4561 caatgggacc gataacgaag tagttaccgt gaccgatgag aacactggtg aaatctctga
     4621 gaaagtcaag ctgggcacta aggcactggc tggtcaatgg ctggcttacg gtgttactcg
     4681 cagtgtgact aagcgttcag tcatgacgct ggcttacggg tccaaagagt tcggcttccg
     4741 tcaacaagtg ctggaagata ccattcagcc agctattgat tccggcaagg gtctgatgtt
     4801 cactcagccg aatcaggctg ctggatacat ggctaagctg atttgggaat ctgtgagcgt
     4861 gacggtggta gctgcggttg aagcaatgaa ctggcttaag tctgctgcta agctgctggc
     4921 tgctgaggtc aaagataaga agactggaga gattcttcgc aagcgttgcg ctgtgcattg
     4981 ggtaactcct gatggtttcc ctgtgtggca ggaatacaag aagcctatta aaacgcgcgt
     5041 gcatattatg ttcctcggtc agttcgaaat gcagcctacc attaacacca acaaagatag
     5101 cgagattgat gcacgcaaac aggagtctgg tatcgctcct aactttgtac acagccaaga
     5161 cggtagccac cttcgtaaga ctgtagtgtg ggcacacgag aagtacggaa tcgaatcttt
     5221 tgcactgatt cacgactcct tcggtaccat tccggctgac gctgcgaacc tgttcaaagc
     5281 agtgcgcgaa actatggttg acacatatga gtcttgtgat gtactggctg atttctacga
     5341 ccagttcgct gaccagttgc acgagtctca attggacaaa atgccagcac ttccggctaa
     5401 aggtaacttg aacctccgtg acatcttaga gtcggacttc gcgttcgcgt aaacttaatt
     5461 aacggcactc ctcagcaaat ataatgaccc tcttgataac ccaagagggc attttttaat
     5521 gcccatggcg tttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5581 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5641 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5701 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5761 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5821 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5881 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5941 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6001 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6061 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6121 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6181 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6241 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6301 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6361 ataagggcga cacggaaatg ttgaatactc atactcttcc ttttt
//
