LOCUS       sequence-47102-e        4231 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 4231)
  AUTHORS   Young CL, Raden DL, Caplan JL, Czymmek KJ, Robinson AS
  TITLE     Cassette series designed for live-cell imaging of proteins and 
            high-resolution techniques in yeast.
  JOURNAL   Yeast. 2012 Mar;29(3-4):119-36. doi: 10.1002/yea.2895. Epub 2012 Apr
            4.
  PUBMED    22473760
REFERENCE   2  (bases 1 to 4231)
  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: "Yeast."; 
            date: "2012-03"; volume: "29(3-4):119-36. doi"; pages: " 
            10.1002/yea.2895. Epub 2012 Apr 4"
COMMENT     Sequence Label: pCY 3090-07
FEATURES             Location/Qualifiers
     source          1..4231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             87..797
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(232..250)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     500..519
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     gene            851..1772
                     /label=bleMX6
                     /note="yeast selectable marker conferring phleomycin 
                     resistance"
     promoter        851..1194
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1195..1569
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      1575..1772
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1868..1887)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1869..1887)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1974..1991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2145..2733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2225..2244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2904..3695,3696..3764))
                     /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     3527..3546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3765..3869)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3937..3955
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3993..4015)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4115..4134
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4215..4231,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4215..4231,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggcttaaga
       61 tcggtgacgg tgctggttta attaacatgg tgagcaaggg cgaggaggat aacatggcca
      121 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
      181 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
      241 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
      301 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
      361 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
      421 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
      481 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
      541 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
      601 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
      661 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
      721 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
      781 acgagctgta caagtagggc gcgccagatc tgtttagctt gcctcgtccc cgccgggtca
      841 cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc
      901 aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt
      961 tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca
     1021 gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc
     1081 cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc
     1141 ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac aaccatggcc
     1201 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
     1261 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
     1321 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
     1381 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
     1441 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
     1501 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
     1561 caggactgat cagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat
     1621 agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt
     1681 ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg
     1741 tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc
     1801 ccgagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg
     1861 ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga
     1921 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1981 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2041 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2101 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2161 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2221 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2281 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     2341 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2401 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2461 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2521 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2581 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2641 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2701 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2761 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2821 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2881 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2941 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3001 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3061 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3121 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3181 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3241 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3301 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3361 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3421 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3481 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3541 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3601 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3661 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3721 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3781 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3841 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3901 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     3961 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4021 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4081 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4141 gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt
     4201 atcatacaca tacgatttag gtgacactat a
//
