LOCUS       sequence-181796-        5714 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Ptrc-YFP subcloned from pAM5080 into NSII-Km vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5714)
  AUTHORS   Cohen SE, Erb ML, Selimkhanov J, Dong G, Hasty J, Pogliano J, Golden
            SS
  TITLE     Dynamic localization of the cyanobacterial circadian clock proteins.
  JOURNAL   Curr Biol. 2014 Aug 18;24(16):1836-44. doi: 
            10.1016/j.cub.2014.07.036. Epub 2014 Aug 7.
  PUBMED    25127213
REFERENCE   2  (bases 1 to 5714)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cub.2014.07.036"; journalName: "Curr Biol"; date: 
            "2014-08-18- 18"; volume: "24"; issue: "16"; pages: "1836-44"
COMMENT     Sequence Label: pAM5087
FEATURES             Location/Qualifiers
     source          1..5714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     494..513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     747..764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    779..919
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(845..864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2053..2868)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2053..2868)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2776..2795
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2860..2879
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3093..4175)
                     /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     4137..4156
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4176..4253)
                     /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        join(4321..4326,4327..4343,4344..4350)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4358..4374
                     /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)."
     CDS             join(4410..4412,4413..4415,4416..5129)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVRFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4455..4476)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4716..4735)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5063..5084
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      5153..5180
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
       61 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      121 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
      181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      781 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
      841 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
      901 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
      961 ttgtctgctc ccgccatccg cttacagaca agctccgcgg gccacgattt gagggacgaa
     1021 tcttcagcgt gactgaagta cctcgcggca agccatttcc cgatgtcttt ttgttggccg
     1081 ccgatcgctt cggggttaat cctacggcct gcgctgtgat cgaagacacc cccttgggag
     1141 tagcggcagg cgtggcggca ggaatgcaag tgtttggcta cgcgggttcc atgcccgctt
     1201 ggcgtctgca agaagccggt gcccatctca tttttgacga tatgcgactg ctgcccagtc
     1261 tgctccaatc gtcgccaaaa gataactcca cagcattgcc caatccctaa cccctgctcg
     1321 cgccgcaact acacactaaa ccgttcctgc gcgatcgctc ttactgttga tggctcgtgc
     1381 ttaaaaacaa tgcaacccta accgtttcag ctggtgattt tcggacgatt tggcttacag
     1441 ggataactga gagtcaacag cctctgtccg tcattgcaca cccatccatg cactggggac
     1501 ttgactcatg ctgaatcaca tttcccttgt ccattgggcg agaggggagg ggaatcttct
     1561 ggactcttca ctaagcggcg atcgcaggtt cttctaccca agcagtggcg atcgcttgat
     1621 tgcagtcttc aatgctggcc tctgcagcca tcgccgccac caaagcatcg taggcgggac
     1681 gttgttgctc cagtaaagtc ttcgcccgta acaatcccca gcgactgcgt aaatccgctt
     1741 cggcaggatt gcgatcgagt tgccgccaca gttgtttcca ctgggcgcga tcgtcagctc
     1801 ccccttccac gttgccgtag accagttgct ctgccgctgc accggccatc aacacctgac
     1861 accactgttc cagcgatcgc tgactgagtt gcccctgtgc ggcttcggct tctagcgcag
     1921 ctgcttggaa ctgcacaccc ccgctagcat cgcgcgcgcg cgcgcggatg ctagccaatg
     1981 gcttattttc ctgctaattg gtaataccat ggtaccgctc tgccagtgtt acaaccaatt
     2041 aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     2101 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     2161 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     2221 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     2281 atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg
     2341 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     2401 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     2461 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     2521 tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag
     2581 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     2641 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     2701 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     2761 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     2821 atttaatcgc ggcctggagc aagacgtttc ccgttgaata tggctcataa caccccttgt
     2881 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     2941 aatgtaacat cagagatttt gagacacaac gtggaccggt atcccgccgc cgccgccgga
     3001 taccggttaa tagtacctgt gaagtgaaaa atggcgcaca ttgtgcgaca tttttttctc
     3061 gagcaggtag tcggccaata acccagggat tttcactgcc cgctttccag tcgggaaacc
     3121 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3181 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     3241 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     3301 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     3361 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     3421 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     3481 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     3541 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     3601 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     3661 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     3721 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     3781 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     3841 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     3901 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     3961 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     4021 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     4081 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     4141 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     4201 tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt gtcaacgtaa
     4261 atgcatgccg cttcgccttc gcgcgcgaat tgatctgcgg cggccgcctg aaatgagctg
     4321 ttgacaatta atcatccggc tcgtataatg tgtggaattg tgagcggata acaatttcac
     4381 acgcatgcgg atcctaagga ggaaaaaaaa tggtgagcaa gggcgaggag ctgttcaccg
     4441 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     4501 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4561 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt
     4621 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4681 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4741 ccgaggtgag gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4801 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4861 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4921 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4981 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
     5041 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     5101 ctctcggcat ggacgagctg tacaagtaaa aatgaaggag ctccccggca ggagaaggcc
     5161 atcctgacgg atggcctttt tgaagctttc tagaatcggg ggcccccggg gggattctag
     5221 agttgctacc aagtagtgac ccgcttcgtg atgcaaaatc cgctgacgat attcgggcga
     5281 tcgctgctga atgccatcga gcagtaacgt ggcaccccgc ccctgccaag tcaccgcatc
     5341 cagactgaac agcaccaaga ggctaaaacc caatcccgcc ggtagcagcg gagaactacc
     5401 cagcattggt cccaccaaag ctaatgccgt cgtggtaaaa atcgcgatcg ccgtcagact
     5461 caagcccagt tcgctcatgc ttcctcatct aggtcacagt cttcggcgat cgcatcgatc
     5521 tgatgctgca gcaagcgttt tccataccgg cgatcgcgcc gtcgcccttt cgctgccgtg
     5581 gcccgcttac gagctcgttt atcgaccacg atcgcatcca aatccgcgat cgcttcccag
     5641 tccggcaatt cagtctgggg cgtccgtttc attaatcctg atagcgtcag accccgtaga
     5701 aaagatcaaa ggat
//
