LOCUS       sequence-12734-e        4649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4649)
  AUTHORS   Tagwerker C, Zhang H, Wang X, Larsen LS, Lathrop RH, Hatfield GW, 
            Auer B, Huang L, Kaiser P
  TITLE     HB tag modules for PCR-based gene tagging and tandem affinity 
            purification in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2006 Jun . 23(8):623-32.
  PUBMED    16823883
REFERENCE   2  (bases 1 to 4649)
  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: "Yeast. 2006
            Jun . 23(8):623-32."
COMMENT     Sequence Label: pFA6a-BIO-hphMX4
FEATURES             Location/Qualifiers
     source          1..4649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      338..525
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(393..417)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            600..2175
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        600..943
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             944..1972
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             944..1972
                     /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      1978..2175
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2286..2305)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2287..2305)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2392..2409)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2563..3151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2643..2662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3322..4113,4114..4182))
                     /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     3945..3964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4183..4287)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4355..4373
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4411..4433)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4533..4552
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4633..4649,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4633..4649,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acgctggaaa ggccggtgaa ggtgctggaa aggccggtga aggtgaaatc cctgcccctc
      121 ttgctggtac cgtttctaag atactggtaa aagaaggtga cactgttaaa gctggtcaaa
      181 cagttctggt gctggaggct atgaaaatgg agacagaaat taacgctcct actgacggaa
      241 aagttgaaaa ggtgttagtt aaggaaagag atgctgttca aggtggtcaa ggtctaatca
      301 agatcggcgt ttaatgaggc gcgccacttc taaataagcg aatttcttat gatttatgat
      361 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
      421 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
      481 tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga
      541 taacagggta atatagatct gtttagcttg ccttgtcccc gccgggtcac ccggccagcg
      601 acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat
      661 gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac
      721 attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag acctgcgagc
      781 agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa
      841 atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct
      901 tgctaggata cagttctcac atcacatccg aacataaaca accatgggta aaaagcctga
      961 actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct
     1021 gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg
     1081 atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg
     1141 gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga
     1201 gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga
     1261 aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc
     1321 cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac
     1381 tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt
     1441 gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc
     1501 cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct
     1561 gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc
     1621 ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca
     1681 gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta
     1741 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     1801 tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg
     1861 gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact
     1921 cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat aatcagtact
     1981 gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta
     2041 gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc
     2101 tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg
     2161 ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagtgtc gagtttaaac
     2221 gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc
     2281 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat
     2341 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2401 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2461 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2521 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2581 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2641 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2701 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     2761 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2821 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2881 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2941 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3001 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3061 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3121 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3181 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3241 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3301 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3361 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3421 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3481 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3541 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3601 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3661 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3721 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3781 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3841 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3901 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3961 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4021 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4081 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4141 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4201 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4261 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4321 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4381 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4441 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4501 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     4561 gcaccatatg gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat
     4621 catacacata cgatttaggt gacactata
//
