LOCUS       sequence-479466-        5111 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Tag S. pombe genes at 3' end (C-terminal end of protein) with 
            superfolder GFP, codon-optimized for S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5111)
  TITLE     Hauf Lab Plasmids 2025
REFERENCE   2  (bases 1 to 5111)
  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"
COMMENT     Sequence Label: pFA6a-psfGFP-hphNT1
FEATURES             Location/Qualifiers
     source          1..5111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             61..771
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      795..982
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(850..874)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1057..1400
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1401..2429
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1401..2429
                     /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      2437..2684
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2453..2471)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2746..2765)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2747..2765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2852..2869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3023..3611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3103..3122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3782..4573,4574..4642))
                     /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     4405..4424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4643..4747)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4815..4833
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4871..4893)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4993..5012
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5093..5111
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5093..5110
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 cgtaagggtg aagaattgtt tactggagtt gttcctattt tagttgaatt ggatggagat
      121 gttaatggtc ataagttttc tgttcgtggt gaaggagaag gagatgctac taacggaaaa
      181 ttgacactta agtttatttg tactacaggt aaacttcctg ttccatggcc tactcttgtt
      241 actacattga catatggtgt tcaatgcttt gctcgttacc cagatcatat gaagcaacat
      301 gatttcttta agtcagcaat gcctgaagga tacgttcaag aacgtactat ttcttttaaa
      361 gatgatggta cttacaagac aagagctgag gttaagtttg aaggagatac tttggttaac
      421 cgaattgaac ttaagggtat tgattttaag gaagatggaa acattcttgg tcataagctt
      481 gaatacaact ttaactcaca taacgtttac attactgctg ataagcaaaa gaatggaatt
      541 aaagcaaact ttaagattcg tcataacgtt gaagatggtt ctgttcaact tgcagatcat
      601 taccaacaaa acactccaat tggagatggt ccagttcttc ttcctgataa ccattatctt
      661 tctacacaat cagttttgtc taaggatcct aacgaaaaga gagatcatat ggttttatta
      721 gaatttgtta ctgctgctgg tattacacat ggtatggatg aactttacaa gtagggcgcg
      781 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
      841 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
      901 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
      961 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt
     1021 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     1081 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     1141 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     1201 agcaaaaatt acggctcctc gctgcggacc tgcgagcagg gaaacgctcc cctcacagac
     1261 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     1321 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     1381 acatccgaac ataaacaacc atgggtaaaa agcctgaact caccgcgacg tctgtcgaga
     1441 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     1501 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     1561 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     1621 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     1681 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     1741 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     1801 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     1861 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     1921 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     1981 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     2041 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     2101 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     2161 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     2221 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     2281 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     2341 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     2401 ccagcactcg tccgagggca aaggaataat ctcgagtcat gtaattagtt atgtcacgct
     2461 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     2521 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     2581 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     2641 ccttgcttga gaaggttttg ggacgctcga aggctttaac gagctcgaat tcatcgatga
     2701 tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg
     2761 tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag
     2821 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2881 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2941 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3001 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3061 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3421 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     3721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3781 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3841 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3901 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3961 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4021 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4081 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4141 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4201 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4261 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4321 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4381 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4441 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4501 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4561 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4621 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4681 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4741 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     4801 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     4861 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     4921 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     4981 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt
     5041 cgttagaacg cggctacaat taatacataa ccttatgtat catacacata cgatttaggt
     5101 gacactatag a
//
