LOCUS       sequence-257588-        4608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression plasmid used in the growth selections of 
            AmiE I122L..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4608)
  AUTHORS   Faber MS, Wrenbeck EE, Azouz LR, Steiner PJ, Whitehead TA
  TITLE     Impact of In Vivo Protein Folding Probability on Local Fitness 
            Landscapes.
  JOURNAL   Mol Biol Evol. 2019 Dec 1;36(12):2764-2777. doi: 
            10.1093/molbev/msz184.
  PUBMED    31400199
REFERENCE   2  (bases 1 to 4608)
  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"; journalName: 
            "Mol Biol Evol"; date: "2019-12-1- 1"; volume: "36"; issue: "12"; 
            pages: "2764-2777"
COMMENT     Sequence Label: AmiE_I122L_pAG
FEATURES             Location/Qualifiers
     source          1..4608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     819..836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    851..990
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(916..935)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1076..1098
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1092..1283)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             2988..3005
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3058..3076)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3072..3119
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3156..3611
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3243..3262)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3453..3474
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3703..4494,4495..4563))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4326..4345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac ctcgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      841 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      901 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
      961 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     1021 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     1081 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     1141 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     1201 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     1261 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     1321 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     1381 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     1441 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     1501 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     1561 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     1621 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     1681 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     1741 cgcgctgagg acccgtgggg ccgccatgcc ggccacagct aacaccacgt cgtccctatc
     1801 tgctgcccta ggtctatgag tggttgctgg ataacttggc gggcatgcat aaggctcgta
     1861 caatatattc agggagacca caacggtttc cctctacaaa taattttgtt taactttcta
     1921 gaaataattt tgtttaactt taagaagttt ttatacatat gagacatggc gatattagct
     1981 cgtcaaatga taccgtaggc gtagccgtgg tgaattacaa gatgccgcgt ttacatactg
     2041 ctgctgaagt cctggataat gcccgcaaaa ttgcggaaat gatcgttggt atgaagcaag
     2101 gtctgccggg catggatctg gttgtgtttc ctgaatattc tttacagggt attatgtacg
     2161 accctgctga aatgatggaa acagccgtgg cgattccagg cgaagaaacg gaaatcttta
     2221 gccgtgcttg tagaaaagca aatgtttggg gtgtgttctc cctgaccggc gaacgtcatg
     2281 aagaacaccc tagaaaggca ccatacaaca ctctggtctt gctcgataac aacggtgaaa
     2341 tcgtacaaaa gtacagaaag atcatcccat ggtgtccgat tgaaggctgg tatccaggtg
     2401 gccagacata cgtctctgaa ggtccgaaag gcatgaagat ctcattaatt atctgcgatg
     2461 acggtaatta tccggaaatt tggagagatt gtgccatgaa gggtgcggaa ttgatcgttc
     2521 gctgccaagg ctatatgtac cctgctaaag accaacaagt tatgatggct aaggcaatgg
     2581 cctgggcgaa taactgttat gtcgctgtag caaacgctgc aggttttgat ggcgtttata
     2641 gctacttcgg tcatagtgcc attatcggtt ttgacggccg tactctgggt gaatgcggcg
     2701 aagaagaaat gggcattcaa tacgcgcagt tgtctctgtc acaaatccgc gatgcccgtg
     2761 cgaatgacca aagtcagaac catttgttta aaatcttgca cagaggttac tccggtttgc
     2821 aggcttcggg cgatggcgac cgtggtctgg cagaatgccc atttgaattc taccgtacct
     2881 gggttactga tgctgaaaag gcaagagaaa acgtggaacg cctgactcgc tccacaacag
     2941 gtgtcgccca atgcccagta ggtcgtctgc cgtatgaagg cctcgagcac caccaccacc
     3001 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     3061 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     3121 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     3181 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3241 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3301 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3361 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3421 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3481 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3541 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3601 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3661 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttaccaat gcttaatcag
     3721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
     4021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4561 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgt
//
