LOCUS       sequence-479470-        5126 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Tag S. pombe genes at 3' end (C-terminal end of protein) with 
            circularly permuted superfolder GFP, codon-optimized for S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  AUTHORS   Kamenz J, Mihaljev T, Kubis A, Legewie S, Hauf S
  TITLE     Robust Ordering of Anaphase Events by Adaptive Thresholds and 
            Competing Degradation Pathways.
  JOURNAL   Mol Cell. 2015 Nov 5;60(3):446-59. doi: 
            10.1016/j.molcel.2015.09.022. Epub 2015 Oct 29.
  PUBMED    26527280
REFERENCE   2  (bases 1 to 5126)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2015.09.022"; journalName: "Mol Cell"; date: 
            "2015-11-5- 5"; volume: "60"; issue: "3"; pages: "446-59"
COMMENT     Sequence Label: pFA6a-psfGFPcp-hphNT1
FEATURES             Location/Qualifiers
     source          1..5126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      810..997
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(865..889)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1072..1415
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1416..2444
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1416..2444
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2452..2699
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2468..2486)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2761..2780)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2762..2780)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2867..2884)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3038..3626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3118..3137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3797..4588,4589..4657))
                     /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     4420..4439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4658..4762)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4830..4848
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4886..4908)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5008..5027
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5108..5126
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5108..5125
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gaagatggtt ctgttcaact tgcagatcat taccaacaaa acactccaat tggagatggt
      121 ccagttcttc ttcctgataa ccattatctt tctacacaat cagttttgtc taaggatcct
      181 aacgaaaaga gagatcatat ggttttatta gaatttgtta ctgctgctgg tattacacat
      241 ggtatggatg aactttacaa gggaggtggc tctatgcgta agggtgaaga attgtttact
      301 ggagttgttc ctattttagt tgaattggat ggagatgtta atggtcataa gttttctgtt
      361 cgtggtgaag gagaaggaga tgctactaac ggaaaattga cacttaagtt tatttgtact
      421 acaggtaaac ttcctgttcc atggcctact cttgttacta cattgacata tggtgttcaa
      481 tgctttgctc gttacccaga tcatatgaag caacatgatt tctttaagtc agcaatgcct
      541 gaaggatacg ttcaagaacg tactatttct tttaaagatg atggtactta caagacaaga
      601 gctgaggtta agtttgaagg agatactttg gttaaccgaa ttgaacttaa gggtattgat
      661 tttaaggaag atggaaacat tcttggtcat aagcttgaat acaactttaa ctcacataac
      721 gtttacatta ctgctgataa gcaaaagaat ggaattaaag caaactttaa gattcgtcat
      781 aacgtttagg gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta
      841 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
      901 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
      961 agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg
     1021 taatatagat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag
     1081 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt
     1141 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat
     1201 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc ggacctgcga gcagggaaac
     1261 gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa
     1321 ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga
     1381 tacagttctc acatcacatc cgaacataaa caaccatggg taaaaagcct gaactcaccg
     1441 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     1501 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     1561 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
     1621 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga
     1681 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
     1741 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
     1801 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
     1861 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     1921 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
     1981 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
     2041 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
     2101 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
     2161 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
     2221 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     2281 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
     2341 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
     2401 gtggaaaccg acgccccagc actcgtccga gggcaaagga ataatctcga gtcatgtaat
     2461 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     2521 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     2581 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     2641 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaacgagct
     2701 cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct
     2761 ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc
     2821 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2881 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2941 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3001 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3061 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3121 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3181 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3241 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3301 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3361 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3421 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3481 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3541 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3601 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3661 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3721 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3901 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4321 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4381 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4441 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4741 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     4801 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     4861 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4921 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4981 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5041 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
     5101 acatacgatt taggtgacac tataga
//
