LOCUS       sequence-252097-        4887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For tagging genes at 3'end (C-terminal end of protein) in S. 
            cerevisiae with mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4887)
  TITLE     mNeonGreen tagging tools for yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4887)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pFA6a-mNeonGreen-KanMX6
FEATURES             Location/Qualifiers
     source          1..4887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..85
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        166..184
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     166..183
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      984..1171
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1039..1063)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1246..2602
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1246..1589
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1590..2399
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1657..1676)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2405..2602
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2706..2725)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2707..2725)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2812..2829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2983..3571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3063..3082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3742..4533,4534..4602))
                     /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     4365..4384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4603..4707)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4775..4793
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4831..4853)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
       61 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga
      121 acgcggctac aattaataca taaccttatg tatcatacac atacgattta ggtgacacta
      181 tagaacgcgg ccgccagctg aagcttcgta cgctgcaggt cgacggatcg gtgacggtgc
      241 tggtttaatt aaatggtgag taaaggcgag gaggacaata tggcgagctt acctgctacc
      301 cacgaactgc acattttcgg ctcaatcaat ggtgtagatt ttgacatggt cggtcagggg
      361 actggtaacc ctaacgatgg ttatgaggag ctaaatctga aatctactaa aggggacctt
      421 caatttagtc cctggatact tgttccgcac ataggttatg gtttccacca atatttgccc
      481 taccctgacg ggatgagccc cttccaggcg gctatggtag atggcagtgg ctatcaggtc
      541 cacagaacaa tgcaatttga ggacggggcc agcctaaccg tgaattatag gtacacgtac
      601 gaggggtcac atataaaggg cgaagcacag gtgaagggga cggggttccc ggcggacgga
      661 ccagtgatga caaactccct gaccgctgcc gattggtgcc gtagtaagaa gacttatcct
      721 aacgacaaaa cgataatcag taccttcaag tggagttata ccacgggcaa cgggaaaaga
      781 taccgtagca cggcgagaac aacttatact ttcgccaagc caatggctgc caactacttg
      841 aagaatcagc ccatgtatgt attccgtaag acggagctga aacatagcaa aaccgaacta
      901 aacttcaagg agtggcagaa agcgtttacc gatgtgatgg gtatggacga attgtataag
      961 taaggcgcgc cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
     1021 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     1081 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg
     1141 aggtcgctct tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat
     1201 agatctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag
     1261 aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg
     1321 tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc
     1381 acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg aaacgctccc
     1441 ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag
     1501 gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt
     1561 tctcacatca catccgaaca taaacaacca tgggtaagga aaagactcac gtttcgaggc
     1621 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     1681 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
     1741 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     1801 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     1861 atgcatggtt actcaccact gcgatccccg gcaaaacagc attccaggta ttagaagaat
     1921 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     1981 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     2041 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     2101 ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag
     2161 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     2221 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2281 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2341 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2401 cagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta
     2461 tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga
     2521 catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat
     2581 gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtttaaac
     2641 gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc
     2701 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat
     2761 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2821 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2881 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3421 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3601 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3661 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3721 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3781 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3841 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3901 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3961 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4021 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4081 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4141 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4201 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4261 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4321 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4381 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4441 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4501 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4561 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4621 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4681 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4741 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4801 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4861 cacagcttgt ctgtaagcgg atgccgg
//
