LOCUS       sequence-351969-        3833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for genome integration expressing constitutively mCherry 
            under pTEFmut8 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3833)
  AUTHORS   Torello Pianale L, Rugbjerg P, Olsson L
  TITLE     Real-Time Monitoring of the Yeast Intracellular State During 
            Bioprocesses With a Toolbox of Biosensors.
  JOURNAL   Front Microbiol. 2022 Jan 7;12:802169. doi: 
            10.3389/fmicb.2021.802169. eCollection 2021.
  PUBMED    35069506
REFERENCE   2  (bases 1 to 3833)
  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: "Front 
            Microbiol."; date: "2022-01-7"; volume: "12:802169. doi"; pages: " 
            10.3389/fmicb.2021.802169. eCollection 2021"
COMMENT     Sequence Label: LT1_33_pTEFmut8-mCherry
FEATURES             Location/Qualifiers
     source          1..3833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        199..596
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             complement(495..506)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             610..1320
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVDGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(755..773)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1023..1042
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1334..1521
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1389..1413)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(join(1966..2757,2758..2826))
                     /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     2589..2608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2827..2931)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3034..3622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3523..3542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagaa cggggtcatc acggctcatc atgcgccaaa caaatgtgtg
       61 caatacacgc tcggatgact gcatgatgac cgcactgact ggggacagca gatccaccta
      121 agcctgtgag agaagcagac acccgacaga tcaaggcagt tacgtctcac caaaccagat
      181 gtcaacacag ctacaacgat agcttcgaaa cgtttctact cctcttttac tcttccagat
      241 cttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      301 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      361 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      421 tttctttttc ttgaaaattc tttttttgat ttttttctct ttcgatgacc tcccatcgat
      481 atttaagtta ataaacggcc ttcaatttcc caagtttcag tcccactttt cttgctctac
      541 tacaactttt tttacctctt gctcattaga aagaaagcat agcaatctaa tctaagtttt
      601 ctagaaaata tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
      661 cgcttcaagg tgcacatgga gggctccgtg gacggccacg agttcgagat cgagggcgag
      721 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
      781 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
      841 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
      901 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
      961 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     1021 ggccccgtaa tgcagaagaa gactatgggc tgggaggcct cctccgagcg gatgtacccc
     1081 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     1141 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     1201 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     1261 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     1321 atcctaactc gaggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     1381 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     1441 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg aggtcgctct
     1501 tattgaccac acctctaccg gcatgccgag caaatgcctg caaatcgctc cccatttcgc
     1561 tgagcatgag acggaaatct gctcgtcagt ggtgctcaca ctgacgaatc atgtacagat
     1621 cataccgatg actgcctggc gactcacaac taagcaagac agccggaacc agcgccggcg
     1681 aacaccactg catatatggc atatcacaac agtccaacta gtgcactgca gtacataacc
     1741 cctaggtctt cctgagagtg acgaaggcta aggtcacttc ttcgtttcct ttattggggt
     1801 ttccgtgtag ccttcccctg aatagtgtgg gacgttttat gagaagccgt aagaaatagc
     1861 cgagcggccg cgattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2101 accgcgggac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2581 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2941 ccgaaaagtg ccacctgtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3001 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3061 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3661 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3721 tgcggccgcc aatgtcaaaa gatcctctca taccatatta agtaaattgc ctccatttct
     3781 ttttcctcgg gcagagaaac tcgcaggcaa cttgctctcg aagtggtcac gtg
//
