LOCUS       sequence-218822-        4467 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 4467)
  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 4467)
  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..4467
                     /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(2705..2724)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2791..2882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2883..3692
                     /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(3775..4467,1..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(3794..3813)
                     /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 aaagtgagat caacaatgaa agcaattttc gtactgaaac
     1801 atcttaatca tgctgcggag ggtttctaat gtccctgtcg aaacaagtac tgccacgtga
     1861 tgttcgcatg cgctttcata tggatggctg cgttaatggt catcagttca tcattgaggg
     1921 tgaaggtaca ggcaaaccgt atgagggcaa gaagattctg aaactgcgcg tgacgaaagg
     1981 tggcccgctg ccgtttgcgt tcgatatcct gagctctgtt tttacttacg gtaaccgttg
     2041 tttttgcgat tatccagagg acatgccgga ctactttaaa cagagcctgc cggaaggtta
     2101 ctcttgggaa cgcaccctga tgtttgagga tggcggttgt ggtacggcga gcgcgcacat
     2161 tagtctggat aagaactgct tcattcacaa tagcaccttc catggcgtca atttcccggc
     2221 agacggtccg gtcatgcaaa agaaaacgct gaactgggag ccgagcagcg aactgattac
     2281 ggcgtgcgac ggtatcctga aaggcgatgt gaccatgttt ctgttgctgg aaggtggcca
     2341 ccgtcttaaa tgtcagttca ccaccagcta caaagcccac aaggcagtta agattccgcc
     2401 gaatcacatt atcgaacacg tgcttgttaa aaaagaggtt gccgacggct ttcagatcca
     2461 agagcatgcg gtcgcaaagc acttcaccgt cgacgttaaa gaaacgtaac cccaagggcg
     2521 acacccccta attagcccgg gcgaaaggcc cagtctttcg actgagcctt tcgttttatt
     2581 tgatgcctgg cagttcccta ctctcgcatg gggagtcccc acactaccat cggcgctacg
     2641 gcgtttcact tctgagttcg gcatggggtc aggtgggacc accgcgctac tgccgccagg
     2701 caaacaaggg gtgttatgag ccatattcag gtataaatgg gctcgcgata atgttcagaa
     2761 ttggttaatt ggttgtaaca ctgaccccta tttgtttatt tttctaaata cattcaaata
     2821 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2881 atatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2941 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3001 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3061 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgccacttc
     3121 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3181 ccggaaaaac agcgttccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3241 atgcgctggc agtgttcctg cgccggttgc actcgattcc tgtttgtaat tgtcctttta
     3301 acagcgatcg cgtatttcgc ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     3361 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3421 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3481 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3541 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3601 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3661 aatttcattt gatgctcgat gagtttttct aagcggcgcg ccatcgaatg gcgcaaaacc
     3721 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatatgaaa
     3781 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     3841 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     3901 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     3961 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     4021 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     4081 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     4141 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     4201 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     4261 catgaggacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     4321 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     4381 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     4441 atgtccggtt ttcaacaaac catgcaa
//
