LOCUS       sequence-48798-e        4988 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 4988)
  AUTHORS   Strack RL, Hein B, Bhattacharyya D, Hell SW, Keenan RJ, Glick BS
  TITLE     A rapidly maturing far-red derivative of DsRed-Express2 for 
            whole-cell labeling.
  JOURNAL   Biochemistry. 2009 Sep 8;48(35):8279-81.
  PUBMED    19658435
REFERENCE   2  (bases 1 to 4988)
  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: 
            "Biochemistry."; date: "2009-09-8"; volume: "48(35)"; pages: 
            "8279-81"
COMMENT     Sequence Label: YIPlac204TKC-E2-Crimson-HDEL
FEATURES             Location/Qualifiers
     source          1..4988
                     /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(539..1213)
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(796..815)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      1210..1219
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1370..1889)
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     primer_bind     complement(1958..1977)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2077..2099
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2218..2892)
                     /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(2893..2994)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2996..3014)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3082..3186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3187..3255,3256..4047)
                     /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(3405..3424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4218..4806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4707..4726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4960..4977
                     /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 tctagattac aattcgtcgt gcttgtacaa ttcgtccata ccgtgagtct
      541 ggaacaggtg gtggcgggcc tcggcgcgct cgtactgctc caccacggtg tagtcctcgt
      601 tgtgggaggt gatgtcgagc ttgtagtcca cgtagtggta gccgggcagc ttcacgggct
      661 tcttggccat gtagatggac ttgaactcac acaggtagtg gccgccgccc ttcagcttca
      721 gcgccatgtg gttctcgccc ttcagcacgc cgtcgcgggg gtagttgcgc tcagtggagg
      781 gctcccagcc cagagtcttc ttctgcatta cggggccgtc ggaggggaag ttcacgccga
      841 tgaacttcac gtggtagatg agggtgccgt cctgcaggga ggagtcctgg gtcacggtca
      901 ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggact
      961 gcttgaggta gtcggggatg tcggcggggt gcttgatgta cgccttggag ccgtagaaga
     1021 actgggggga caggatgtcc caggcgaagg gcagggggcc gcccttggtc acttgcagct
     1081 tggcggtctg ggtgccctcg tagggcttgc cctcgcccac gccctcgatc tcgaactcgt
     1141 ggccgttcac ggagccctcc atgtgcacct tgaagcgcat gaagggcttg atgacgttct
     1201 cagtgctatc catggtggcg actagtggat ccccatcatc ggcacctcta actaaaacat
     1261 tggaggagtg gaaagaattc tgtaaaggta atattaccac gaaaagggcg tacaggacca
     1321 cggagagtgg taccagcagc ttgccagcgc ttagtctgtt gaaaaacatt tttagtttat
     1381 gtatgtgttt tttgtagtta tagatttaag caagaaaaga atacaaacaa aaaattgaaa
     1441 aagattgatt tagaattaaa aagaaaaata tttacgtaag aagggaaaat agtaaatgtt
     1501 gcaagttcac taaactccta aattatgctg ccctttatat tccctgttac agcagccgag
     1561 ccaaaggtat ataggctcct ttgcattagc atgcgtaaca aaccacctgt cagtttcaac
     1621 cgaggtggta tccgagagaa ttgtgtgatt gctttaatta atttcggaga atctcacatg
     1681 ccactgaaga ttaaaaactg gatgccagaa aaggggtgtc caggtgtaac atcaatagag
     1741 gaagctgaaa agtcttagaa cgggtaatct tccaccaacc tgatgggttc ctagatataa
     1801 tctcgaaggg aataagtagg gtgataccgc agaagtgtct gaatgtatta aggtcctcac
     1861 agtttaaatc ccgctcacac taacgtaggc gcctgatgcg gtattttctc cttacgcatc
     1921 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     1981 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2041 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2101 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcga tcttttatgc ttgcttttca
     2161 aaaggcttgc aggcaagtgc acaaacaata cttaaataaa tactactcag taataaccta
     2221 tttcttagca tttttgacga aatttgctat tttgttagag tcttttacac catttgtctc
     2281 cacacctccg cttacatcaa caccaataac gccatttaat ctaagcgcat caccaacatt
     2341 ttctggcgtc agtccaccag ctaacataaa atgtaagctc tcggggctct cttgccttcc
     2401 aacccagtca gaaatcgagt tccaatccaa aagttcacct gtcccacctg cttctgaatc
     2461 aaacaaggga ataaacgaat gaggtttctg tgaagctgca ctgagtagta tgttgcagtc
     2521 ttttggaaat acgagtcttt taataactgg caaaccgagg aactcttggt attcttgcca
     2581 cgactcatct ccatgcagtt ggacgatatc aatgccgtaa tcattgacca gagccaaaac
     2641 atcctcctta ggttgattac gaaacacgcc aaccaagtat ttcggagtgc ctgaactatt
     2701 tttatatgct tttacaagac ttgaaatttt ccttgcaata accgggtcaa ttgttctctt
     2761 tctattgggc acacatataa tacccagcaa gtcagcatcg gaatctagtg cacattctgc
     2821 ggcctctgtg ctctgcaagc cgcaaacttt caccaatgga ccagaactac ctgtgaaatt
     2881 aataacagac atactccaag ctgcctttgt gtgcttaatc acgtatactc acgtgctcaa
     2941 tagtcaccaa tgccctccct cttggccctc tccttttctt ttttcgaccg aattggcctc
     3001 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3061 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3301 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3361 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3421 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3481 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3541 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3601 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3661 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3721 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3781 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3841 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3901 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3961 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4021 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4081 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4141 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     4381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4921 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4981 gcggaaga
//
