LOCUS       sequence-270428-        3589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAC6 experimental evolution.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3589)
  AUTHORS   Stiffler MA, Poelwijk FJ, Brock KP, Stein RR, Riesselman A, Teyra J,
            Sidhu SS, Marks DS, Gauthier NP, Sander C
  TITLE     Protein Structure from Experimental Evolution.
  JOURNAL   Cell Syst. 2020 Jan 22;10(1):15-24.e5. doi: 
            10.1016/j.cels.2019.11.008. Epub 2019 Dec 11.
  PUBMED    31838147
REFERENCE   2  (bases 1 to 3589)
  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"; doi: 
            "10.1016/j.cels.2019.11.008"; journalName: "Cell Syst"; date: 
            "2020-01-22- 22"; volume: "10"; issue: "1"; pages: "15-24.e5"
COMMENT     Sequence Label: pBR322_ZA-ampR-PSEAB-2A
FEATURES             Location/Qualifiers
     source          1..3589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..103)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      883..1471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1372..1391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1625..1642
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1657..1797
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1723..1742)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1883..1905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1899..2090)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(2729..3520,3521..3589))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3352..3371
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
       61 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
      121 gatatcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      181 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      241 ataatattga cacaggagac caaaatgatc cgtattcagc cgggagttat agcaaatcca
      301 catcagtggt tacaattacg tcaggcactt tggcctgatt gtccagccga tgagcatcaa
      361 gcggaaatcc gccaaatgct cgcagagcca cagcgttatg gatgttggtt ggctcttgcc
      421 gccgactcac aagcacttgg cctggcggaa gcttcacttc ggcatgatta tgtgaatggc
      481 acagactcat cgcctgtggt ttttttggaa gggatttatg tagcgcctga acatcgtcgt
      541 caaggtatcg cacagcggtt gattgaacaa gtgtcacaat gggggcgaca gttgggatgt
      601 acacagttgg cttcggatac agggttggat aatctggcca gccaggccct gcatcaagca
      661 ctcggttttg ctgaaactga acgtgtggtg tattttaaga aagctctcta actgtagaga
      721 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
      781 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
      841 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
      901 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
      961 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1021 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1081 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1141 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1201 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1261 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1321 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1381 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1441 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     1501 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     1561 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     1621 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     1681 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     1741 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     1801 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     1861 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     1921 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     1981 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     2041 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     2101 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     2161 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     2221 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     2281 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     2341 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     2401 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     2461 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     2521 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     2581 atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg
     2641 gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat
     2701 ccgttagcga ggtgccgccg gcttccattt accaatgctt aatcagtgag gcacctatct
     2761 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2821 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgt gacccacgct
     2881 caccggcacc agatttatca gcaataaacc agccagccgg aagtgccgag cgcagaagtg
     2941 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3001 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgctggc atcgtggtgt
     3061 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3121 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3181 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3241 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3301 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     3361 cgccacatag cagaactttg aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3421 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgg gcacccaact
     3481 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3541 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcat
//
