LOCUS       sequence-11857-e        4812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4812)
  AUTHORS   Strack RL, Strongin DE, Bhattacharyya D, Tao W, Berman A, Broxmeyer 
            HE, Keenan RJ, Glick BS
  TITLE     A noncytotoxic DsRed variant for whole-cell labeling.
  JOURNAL   Nat Methods. 2008 Nov . 5(11):955-7.
  PUBMED    18953349
REFERENCE   2  (bases 1 to 4812)
  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 
            Methods. 2008 Nov . 5(11):955-7."
COMMENT     Sequence Label: YIPlac204TC-NLS-DsRed-Express2
FEATURES             Location/Qualifiers
     source          1..4812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(239..486)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     452..470
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(503..1180)
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(565..588)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(763..782)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     980..1000
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     CDS             complement(1193..1213)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1228..1658)
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     primer_bind     complement(1782..1801)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1901..1923
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2042..2716)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2717..2818)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2820..2838)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2906..3010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3011..3079,3080..3871)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3229..3248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4042..4630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4531..4550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4784..4801
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac
      301 atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta
      361 atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt
      421 ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac ataactaatt
      481 acatgatgac tctagaggat ccctactgga acaggtggtg gcgggcctcg gcgcgctcgt
      541 actgctccac cacggtgtag tcctcgttgt gggaggtgat gtccagcttg gagtccacgt
      601 agtagtagcc gggcagcttc acgggcttct tggccatgta gattgacttg aactccacca
      661 ggtagtggcc gccgcccttc agcttcagcg ccttgtggat ctcgcccttc agcacgccgt
      721 cgcgggggta caggcgctcg gtggagggct cccagcccag agtcttcttc tgcattacgg
      781 ggccgtcgga ggggaagttc acgccgatga acttcacgtg gtagatgaag gtgccgtcct
      841 gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc atcacgcgct
      901 cccacttgaa gccctcgggg aaggacagct tcttgtagtc ggggatgtcg gcggggtgct
      961 tcacgtacac cttggagccg tactggaact ggggggacag gatgtcccag gcgaagggca
     1021 gggggccgcc cttggtcacc tgcagcttgg cggtctgggt gccctcgtag ggcttgccct
     1081 cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg tgcaccttga
     1141 agcgcatgaa gggcttgatg acgttctcag tgctatccat ttttgaattg ccgacctttc
     1201 ttttcttttt tggcattttt tgaattcttt tagtttatgt atgtgttttt tgtagttata
     1261 gatttaagca agaaaagaat acaaacaaaa aattgaaaaa gattgattta gaattaaaaa
     1321 gaaaaatatt tacgtaagaa gggaaaatag taaatgttgc aagttcacta aactcctaaa
     1381 ttatgctgcc ctttatattc cctgttacag cagccgagcc aaaggtatat aggctccttt
     1441 gcattagcat gcgtaacaaa ccacctgtca gtttcaaccg aggtggtatc cgagagaatt
     1501 gtgtgattgc tttaattaat ttcggagaat ctcacatgcc actgaagatt aaaaactgga
     1561 tgccagaaaa ggggtgtcca ggtgtaacat caatagagga agctgaaaag tcttagaacg
     1621 ggtaatcttc caccaacctg atgggttcct agatataatc tcgaagggaa taagtagggt
     1681 gataccgcag aagtgtctga atgtattaag gtccgcctga tgcggtattt tctccttacg
     1741 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     1801 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     1861 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     1921 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcgatctttt atgcttgctt
     1981 ttcaaaaggc ttgcaggcaa gtgcacaaac aatacttaaa taaatactac tcagtaataa
     2041 cctatttctt agcatttttg acgaaatttg ctattttgtt agagtctttt acaccatttg
     2101 tctccacacc tccgcttaca tcaacaccaa taacgccatt taatctaagc gcatcaccaa
     2161 cattttctgg cgtcagtcca ccagctaaca taaaatgtaa gctctcgggg ctctcttgcc
     2221 ttccaaccca gtcagaaatc gagttccaat ccaaaagttc acctgtccca cctgcttctg
     2281 aatcaaacaa gggaataaac gaatgaggtt tctgtgaagc tgcactgagt agtatgttgc
     2341 agtcttttgg aaatacgagt cttttaataa ctggcaaacc gaggaactct tggtattctt
     2401 gccacgactc atctccatgc agttggacga tatcaatgcc gtaatcattg accagagcca
     2461 aaacatcctc cttaggttga ttacgaaaca cgccaaccaa gtatttcgga gtgcctgaac
     2521 tatttttata tgcttttaca agacttgaaa ttttccttgc aataaccggg tcaattgttc
     2581 tctttctatt gggcacacat ataataccca gcaagtcagc atcggaatct agtgcacatt
     2641 ctgcggcctc tgtgctctgc aagccgcaaa ctttcaccaa tggaccagaa ctacctgtga
     2701 aattaataac agacatactc caagctgcct ttgtgtgctt aatcacgtat actcacgtgc
     2761 tcaatagtca ccaatgccct ccctcttggc cctctccttt tcttttttcg accgaattgg
     2821 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2881 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2941 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3001 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3061 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3121 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3181 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3241 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3301 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3361 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3421 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3481 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3541 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3601 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3661 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3721 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3781 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3841 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3901 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3961 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4021 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     4201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4681 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4741 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4801 ggaagcggaa ga
//
