LOCUS       sequence-363370-        6276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human HA-tagged DIPP3B from yeast GPD promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6276)
  TITLE     Downey Lab Plasmids
REFERENCE   2  (bases 1 to 6276)
  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"
COMMENT     Sequence Label: p416-GPD-DIPP3B-HA
FEATURES             Location/Qualifiers
     source          1..6276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    69..572
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(714..782,783..1574)
                     /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     complement(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2657..2662,2663..2680,2681..2687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2711
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2700..2722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2719..2735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2754..2774
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2756..2774
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2799..3442
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3336..3359
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3448..3464
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3448..3464
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3958..3984
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4008..4024)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4009..4025)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4027..4274
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4043..4061)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4292..4311)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4293..4311)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4321..4338)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4321..4337)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4330..4352)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4478..4933
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4565..4584)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4775..4796
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5064..5867)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5868..6083)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6107..6126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6226..6248
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
      121 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
      181 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
      241 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
      301 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
      361 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
      421 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
      481 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
      541 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     2761 accctcacta aagggaacaa aagctggagc tcagtttatc attatcaata ctcgccattt
     2821 caaagaatac gtaaataatt aatagtagtg attttcctaa ctttatttag tcaaaaaatt
     2881 agccttttaa ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat
     2941 atataacatc gtaggtgtct gggtgaacag tttattcctg gcatccacta aatataatgg
     3001 agcccgcttt ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa aatattgttt
     3061 tcttcaccaa ccatcagttc ataggtccat tctcttagcg caactacaga gaacaggggc
     3121 acaaacaggc aaaaaacggg cacaacctca atggagtgat gcaacctgcc tggagtaaat
     3181 gatgacacaa ggcaattgac ccacgcatgt atctatctca ttttcttaca ccttctatta
     3241 ccttctgctc tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa
     3301 attattcccc tacttgacta ataagtatat aaagacggta ggtattgatt gtaattctgt
     3361 aaatctattt cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta
     3421 aaacaccaga acttagtttc gacggattct agaactagtg gatccatgaa gtgcaaaccc
     3481 aaccagacgc ggacctacga ccccgagggg ttcaagaagc gggcggcgtg cctgtgcttc
     3541 cggagcgaac gcgaggacga ggtcctgtta gtgagtagca gccggtaccc ggaccgctgg
     3601 atcgtgccgg gcgggggcat ggagcccgag gaggagccgg gcggtgcggc ggtccgagag
     3661 gtgtacgaag aagcgggagt caaggggaag ttaggccggc tcctgggcgt cttcgaacag
     3721 aaccaggatc gcaagcacag aacgtacgtg tatgtactga ctgtcacgga gctgctggag
     3781 gattgggaag attcggttag cattgggagg aagcgagagt ggttcaaagt cgaagatgcc
     3841 atcaaggttc tccagtgcca caagcccgtg cacgccgaat atctggagaa actaaagctg
     3901 ggcggttccc caaccaatgg aaactccatg gccccatcct cgccagatag cgatccctac
     3961 ccatacgatg ttccagatta cgcttaagaa ttcgatatca agcttatcga taccgtcgac
     4021 ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc
     4081 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag
     4141 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac
     4201 gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga
     4261 aggctttaat ttgcggccgg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     4321 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4381 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4441 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta
     4501 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4561 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4621 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4681 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4741 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4801 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4861 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4921 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4981 gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat
     5041 gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc
     5101 cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt
     5161 cctcttccaa caataataat gtcagatcct gtagagacca catcatccac ggttctatac
     5221 tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa
     5281 tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct
     5341 ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc
     5401 ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc
     5461 gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct
     5521 gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg
     5581 tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc
     5641 ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa
     5701 caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc
     5761 aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca
     5821 ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt
     5881 ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct
     5941 tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc
     6001 ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa
     6061 taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca atctgctctg
     6121 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     6181 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     6241 gtcagaggtt ttcaccgtca tcaccgaaac gcgcga
//
