LOCUS       sequence-90572-e        4610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RFP from coral Corynactis sp. under T5 promoter with lac operators; 
            kanamycin resistant..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4610)
  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 4610)
  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: p005kanRFP
FEATURES             Location/Qualifiers
     source          1..4610
                     /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"
     protein_bind    1807..1823
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     promoter        join(1848..1862,1863..1868,1869..1885,1886..1892)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1868..1884
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     protein_bind    1898..1917
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by 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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     RBS             1958..1966
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(2848..2867)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        2934..3025
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3026..3835
                     /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(3918..4610,1..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(3937..3956)
                     /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 aatcagtttt tgatatcaaa attatacatg tcaacgataa tacaaaatat
     1741 aatacaaact ataagatgtt atcagtattt attatgcatt tagaataaat tttgtgtcgc
     1801 ccttaattgt gagcggataa caattacgag cttcatgcac agtgaaatca tgaaaaattt
     1861 atttgctttg tgagcggata acaattataa tatgtggaat tgtgagcgct cacaattcca
     1921 caacggtttc cctctagaaa taattttgtt taacttttaa ggaggtaaaa aatgtccctg
     1981 tcgaaacaag tactgccacg tgatgttcgc atgcgctttc atatggatgg ctgcgttaat
     2041 ggtcatcagt tcatcattga gggtgaaggt acaggcaaac cgtatgaggg caagaagatt
     2101 ctgaaactgc gcgtgacgaa aggtggcccg ctgccgtttg cgttcgatat cctgagctct
     2161 gtttttactt acggtaaccg ttgtttttgc gattatccag aggacatgcc ggactacttt
     2221 aaacagagcc tgccggaagg ttactcttgg gaacgcaccc tgatgtttga ggatggcggt
     2281 tgtggtacgg cgagcgcgca cattagtctg gataagaact gcttcattca caatagcacc
     2341 ttccatggcg tcaatttccc ggcagacggt ccggtcatgc aaaagaaaac gctgaactgg
     2401 gagccgagca gcgaactgat tacggcgtgc gacggtatcc tgaaaggcga tgtgaccatg
     2461 tttctgttgc tggaaggtgg ccaccgtctt aaatgtcagt tcaccaccag ctacaaagcc
     2521 cacaaggcag ttaagattcc gccgaatcac attatcgaac acgtgcttgt taaaaaagag
     2581 gttgccgacg gctttcagat ccaagagcat gcggtcgcaa agcacttcac cgtcgacgtt
     2641 aaagaaacgt aaccccaagg gcgacacccc ctaattagcc cgggcgaaag gcccagtctt
     2701 tcgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc atggggagtc
     2761 cccacactac catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg
     2821 accaccgcgc tactgccgcc aggcaaacaa ggggtgttat gagccatatt caggtataaa
     2881 tgggctcgcg ataatgttca gaattggtta attggttgta acactgaccc ctatttgttt
     2941 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3001 tcaataatat tgaaaaagga agaatatgag ccatattcaa cgggaaacgt cgaggccgcg
     3061 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     3121 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     3181 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     3241 gctgacggaa tttatgccac ttccgaccat caagcatttt atccgtactc ctgatgatgc
     3301 atggttactc accactgcga tccccggaaa aacagcgttc caggtattag aagaatatcc
     3361 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcactcgat
     3421 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgcctcgctc aggcgcaatc
     3481 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3541 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3601 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3661 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3721 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3781 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagcggc
     3841 gcgccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     3901 aattcagggt ggtgaatatg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     3961 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     4021 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     4081 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     4141 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     4201 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     4261 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     4321 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     4381 ccatcaacag tattattttc tcccatgagg acggtacgcg actgggcgtg gagcatctgg
     4441 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     4501 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     4561 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
//
