LOCUS       sequence-212479-        4466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synthetic RFP based on Corynactis spp under weak synthetic 
            constitutive promoter pFAB4024; kanamycin resistant..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4466)
  AUTHORS   Siddall A, Williams AA, Sanders J, Denton JA, Madden D, Schollar J, 
            Bryk J
  TITLE     UNIGEMS: plasmids and parts to facilitate teaching on assembly, gene
            expression control and logic in E. coli
  JOURNAL   bioRxiv 2021.06.20.449138
REFERENCE   2  (bases 1 to 4466)
  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 
            2021.06.20.449138"
COMMENT     Sequence Label: p005-RFP-weak
FEATURES             Location/Qualifiers
     source          1..4466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      476..1064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     965..984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1218..1235
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1333..1350
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1334..1350
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2704..2723)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2790..2881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2882..3691
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(3774..4466,1..390)
                     /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     complement(3793..3812)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
       61 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
      121 ggatacgacg ataccgaaga tagctcatgt tatatcccgc cgttaaccac catcaaacag
      181 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
      241 gcggtgaagg gcaatcagct gttgccagtc tcactggtga aaagaaaaac caccctggcg
      301 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      361 caggtttccc gactggaaag cgggcagtga ctcatgacca aaatccctta acgtgagtta
      421 cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      481 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      541 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      601 gagcgcagat accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga
      661 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      721 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      781 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      841 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      901 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      961 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1021 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1081 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     1141 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     1201 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac
     1261 tgccaggcat caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa
     1321 ctctttctgt gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga
     1381 taacgagtaa tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg
     1441 gggggagttt agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg
     1501 atatatgaga attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata
     1561 cgttgctttt tcgattgatg aacacctata attaaactat tcatctatta tttatgattt
     1621 tttgtatata caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata
     1681 ataaagggaa aatcagaaaa agagtattga cttcgcatct ttttgtacct ataatgtgtg
     1741 gagggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg tactgaaaca
     1801 tcttaatcat gctgcggagg gtttctaatg tccctgtcga aacaagtact gccacgtgat
     1861 gttcgcatgc gctttcatat ggatggctgc gttaatggtc atcagttcat cattgagggt
     1921 gaaggtacag gcaaaccgta tgagggcaag aagattctga aactgcgcgt gacgaaaggt
     1981 ggcccgctgc cgtttgcgtt cgatatcctg agctctgttt ttacttacgg taaccgttgt
     2041 ttttgcgatt atccagagga catgccggac tactttaaac agagcctgcc ggaaggttac
     2101 tcttgggaac gcaccctgat gtttgaggat ggcggttgtg gtacggcgag cgcgcacatt
     2161 agtctggata agaactgctt cattcacaat agcaccttcc atggcgtcaa tttcccggca
     2221 gacggtccgg tcatgcaaaa gaaaacgctg aactgggagc cgagcagcga actgattacg
     2281 gcgtgcgacg gtatcctgaa aggcgatgtg accatgtttc tgttgctgga aggtggccac
     2341 cgtcttaaat gtcagttcac caccagctac aaagcccaca aggcagttaa gattccgccg
     2401 aatcacatta tcgaacacgt gcttgttaaa aaagaggttg ccgacggctt tcagatccaa
     2461 gagcatgcgg tcgcaaagca cttcaccgtc gacgttaaag aaacgtaacc ccaagggcga
     2521 caccccctaa ttagcccggg cgaaaggccc agtctttcga ctgagccttt cgttttattt
     2581 gatgcctggc agttccctac tctcgcatgg ggagtcccca cactaccatc ggcgctacgg
     2641 cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact gccgccaggc
     2701 aaacaagggg tgttatgagc catattcagg tataaatggg ctcgcgataa tgttcagaat
     2761 tggttaattg gttgtaacac tgacccctat ttgtttattt ttctaaatac attcaaatat
     2821 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa
     2881 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2941 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     3001 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3061 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgccacttcc
     3121 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3181 cggaaaaaca gcgttccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     3241 tgcgctggca gtgttcctgc gccggttgca ctcgattcct gtttgtaatt gtccttttaa
     3301 cagcgatcgc gtatttcgcc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3361 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3421 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3481 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3541 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3601 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3661 gtttcatttg atgctcgatg agtttttcta agcggcgcgc catcgaatgg cgcaaaacct
     3721 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatatgaaac
     3781 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     3841 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     3901 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     3961 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     4021 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     4081 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     4141 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     4201 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     4261 atgaggacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     4321 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     4381 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     4441 tgtccggttt tcaacaaacc atgcaa
//
