LOCUS       sequence-381135-        5720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the RamR transcription factor in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  AUTHORS   d'Oelsnitz S, Kim W, Burkholder NT, Javanmardi K, Thyer R, Zhang Y, 
            Alper HS, Ellington AD
  TITLE     Using fungible biosensors to evolve improved alkaloid biosyntheses.
  JOURNAL   Nat Chem Biol. 2022 Jul 7. pii: 10.1038/s41589-022-01072-w. doi: 
            10.1038/s41589-022-01072-w.
  PUBMED    35799063
REFERENCE   2  (bases 1 to 5720)
  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 Chem 
            Biol. 2022 Jul 7. pii: 10.1038/s41589-022-01072-w. doi: 
            10.1038/s41589-022-01072-w."
COMMENT     Sequence Label: pReg-RamR
FEATURES             Location/Qualifiers
     source          1..5720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(612..683)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(871..1953)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1915..1934
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1954..2031)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        2309..2343
                     /label=BBa_J23112
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0"
     promoter        2309..2343
                     /label=BBa_J23103
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     CDS             4099..4290
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4284..4306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4447..4466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4541..4558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4712..5300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4792..4811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5471..5720,1..542,543..611))
                     /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"
ORIGIN
        1 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
       61 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      121 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      181 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      241 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      301 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      361 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      421 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      481 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      541 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
      601 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      661 atgagcggat acatatttga atgtatttag aaaaatatgc gccttgagcg acacgaatta
      721 tgcagtgatt tacgacctgc acagccatac cacagcttcc gatggctgcc tgacgccaga
      781 agcattggtg caccgtgcag tcgatgataa gctgtcaaac atgagaattg tgcctaatga
      841 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
      901 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
      961 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     1021 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     1081 ctgtttgatg gtggttaacg gcgggatata acatgagcta tcttcggtat cgtcgtatcc
     1141 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     1201 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     1261 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     1321 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     1381 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     1441 cacgcccagt cgcgtaccat cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     1501 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     1561 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     1621 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     1681 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     1741 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     1801 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     1861 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     1921 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     1981 cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt cgggacgtca
     2041 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gttgcggccg cgaagacagc
     2101 gttatcagag atgagacacc gtgcgataat gtcgggcaat caggtgcgac tcggtaccaa
     2161 attccagaaa agaggggagc gggaaaccgc tccccttttt tcgttttggt cccaatctat
     2221 cgattgtatg gacttcatct tcattacctc gtatcattgt acacctgccg aacgcaaggg
     2281 catgggctgt cgacctttga aaagtaccct gatagctagc tcagtcctag ggattatgct
     2341 agcaattacg agccccatag ggtggtgtgt accacccctg atgagtccaa aaggacgaaa
     2401 tggggcctct acaaataatt ttgtttaacg gaaccacgta tcagaaggag gttagtatat
     2461 ggttgctcgc ccaaagtctg aggacaaaaa gcaggcattg cttgaagcgg caactcaagc
     2521 catcgcgcaa tcaggcattg ccgctagtac cgctgtaatt gcacgcaatg cgggagttgc
     2581 ggaagggacg ttgttccgct atttcgcaac gaaagatgag ttgatcaaca ccctttactt
     2641 acatttgaaa caggacctgt gccaatcaat gatcatggaa ttggatcgtt ctattactga
     2701 cgctaagatg atgacccgtt ttatctggaa cagttatatt agctggggat tgaaccaccc
     2761 agctcgccat cgtgccattc gtcagttggc ggtttctgaa aagttgacga aggaaaccga
     2821 acaacgcgcg gatgatatgt tcccggagtt acgcgacttg tgccaccgta gtgttcttat
     2881 ggtgtttatg tccgacgagt accgcgcctt cggcgacggg ttgttcttgg cgcttgctga
     2941 gacgactatg gatttcgctg cgcgcgaccc ggctcgcgct ggtgagtaca ttgcgttggg
     3001 cttcgaggct atgtggcgcg cacttacgcg cgaagagcag taataatcct aagaattcga
     3061 ggagtgcagg ctcggtaaca tacggtctag ctatctgact atcgccgctg tgagctcggt
     3121 accaaattcc agaaaagagg ccgcgaaagc ggcctttttt cgttttggtc caagcccgat
     3181 gcgccagagt tgtttctgaa acatggcaaa ggtatcacta gtcttcgcgg ccgccatgct
     3241 gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac
     3301 cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg cctcggcgag
     3361 cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc
     3421 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     3481 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     3541 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     3601 cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     3661 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     3721 cgcgagcgaa cgtgaagtga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     3781 atggtcatcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     3841 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     3901 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     3961 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     4021 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     4081 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     4141 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     4201 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     4261 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4321 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4381 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     4441 actatgcggc atcagagcag attgtactga gagtgcaccg gtgtgaaata ccgcacagat
     4501 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     4561 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4621 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4681 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4741 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4801 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4861 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4921 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4981 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5041 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5101 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     5161 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5221 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5281 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5341 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5401 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5461 ggtctaacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5521 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5581 catctggccc cagtgctgca atgataccgc gcgatccacg ctcaccggct ccagatttat
     5641 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5701 cctccatcca gtctattaat
//
