LOCUS       sequence-479480-        5105 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Tag S. pombe genes at 3' end (C-terminal end of protein) with 
            mNeonGreen, codon-optimized for S. pombe.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5105)
  TITLE     Hauf Lab Plasmids 2025
REFERENCE   2  (bases 1 to 5105)
  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-pmNeonGreen-hphNT1
FEATURES             Location/Qualifiers
     source          1..5105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      789..976
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(844..868)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1051..1394
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1395..2423
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1395..2423
                     /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      2431..2678
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2447..2465)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2740..2759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2741..2759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2846..2863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3017..3605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3097..3116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3776..4567,4568..4636))
                     /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     4399..4418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4637..4741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4809..4827
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4865..4887)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4987..5006
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5087..5105
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5087..5104
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
       61 gtttctaagg gtgaagaaga taacatggca tcattgcctg ctacacatga attgcatatt
      121 tttggttcta ttaacggagt tgattttgat atggttggtc aaggaactgg taaccctaac
      181 gatggatacg aagaacttaa ccttaagtct acaaagggag atcttcaatt ttcaccttgg
      241 attcttgttc cacatattgg atacggtttt catcaatact taccttatcc agatggaatg
      301 tctccatttc aagctgcaat ggttgatgga tcaggttatc aagttcatcg aactatgcaa
      361 tttgaagatg gtgcttctct tactgttaac taccgttaca catacgaagg atcacatatt
      421 aaaggtgaag ctcaagttaa gggaactggt tttcctgctg atggtccagt tatgactaat
      481 tctttgacag ctgcagattg gtgtcgttct aagaaaactt atcctaacga taagactatt
      541 atttctactt ttaaatggtc atacactaca ggaaatggta aacgttatag atctactgct
      601 agaactacat acacatttgc aaaacctatg gctgcaaact acttaaagaa ccaaccaatg
      661 tacgtttttc gaaagactga acttaagcat tctaagacag aattgaactt taaggaatgg
      721 caaaaggctt ttactgatgt tatgggtatg gatgaacttt acaaataggg cgcgccactt
      781 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
      841 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
      901 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
      961 accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc tgtttagctt
     1021 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
     1081 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     1141 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     1201 aattacggct cctcgctgcg gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     1261 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     1321 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     1381 gaacataaac aaccatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     1441 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     1501 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     1561 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     1621 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     1681 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     1741 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     1801 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     1861 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     1921 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     1981 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     2041 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     2101 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     2161 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     2221 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     2281 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     2341 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     2401 ctcgtccgag ggcaaaggaa taatctcgag tcatgtaatt agttatgtca cgcttacatt
     2461 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     2521 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     2581 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     2641 ttgagaaggt tttgggacgc tcgaaggctt taacgagctc gaattcatcg atgatatcag
     2701 atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa
     2761 tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2821 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3001 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3061 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3121 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3181 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3241 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3301 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3361 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3421 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3481 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3541 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3601 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3661 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3721 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3781 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3841 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3901 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3961 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4021 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4081 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4141 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4201 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4261 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4321 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4381 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4441 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4501 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4561 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4621 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4681 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4741 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4801 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     4861 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4921 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     4981 actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag
     5041 aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact
     5101 ataga
//
