LOCUS       sequence-8455-ex        5967 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 5967)
  TITLE     Buchler lab plasmids
REFERENCE   2  (bases 1 to 5967)
  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: p404ADH1
FEATURES             Location/Qualifiers
     source          1..5967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2241..2261
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3030..3734
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     3748..3764
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3748..3764
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3798..3814)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3799..3815)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3817..4064
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3833..3851)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4082..4101)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4083..4101)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4111..4128)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4111..4127)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4120..4142)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4272..4727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4359..4378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4569..4590
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4826..5500)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(5501..5781)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5798..5817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5917..5939
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcg
     2281 ccgggatcga agaaatgatg gtaaatgaaa taggaaatca aggagcatga aggcaaaaga
     2341 caaatataag ggtcgaacga aaaataaagt gaaaagtgtt gatatgatgt atttggcttt
     2401 gcggcgccga aaaaacgagt ttacgcaatt gcacaatcat gctgactctg tggcggaccc
     2461 gcgctcttgc cggcccggcg ataacgctgg gcgtgaggct gtgcccggcg gagttttttg
     2521 cgcctgcatt ttccaaggtt taccctgcgc taaggggcga gattggagaa gcaataagaa
     2581 tgccggttgg ggttgcgatg atgacgacca cgacaactgg tgtcattatt taagttgccg
     2641 aaagaacctg agtgcatttg caacatgagt atactagaag aatgagccaa gacttgcgag
     2701 acgcgagttt gccggtggtg cgaacaatag agcgaccatg accttgaagg tgagacgcgc
     2761 ataaccgcta gagtactttg aagaggaaac agcaataggg ttgctaccag tataaataga
     2821 caggtacata caacactgga aatggttgtc tgtttgagta cgctttcaat tcatttgggt
     2881 gtgcacttta ttatgttaca atatggaagg gaactttaca cttctcctat gcacatatat
     2941 taattaaagt ccaatgctag tagagaaggg gggtaacacc cctccgcgct cttttccgat
     3001 ttttttctaa accgtggaat atttcggata tccttttgtt gtttccgggt gtacaatatg
     3061 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
     3121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
     3181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
     3241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
     3301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
     3361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
     3421 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
     3481 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
     3541 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
     3601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
     3661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
     3721 gcatacaatc aactccaagc tggccgctct agaactagtg gatcccccgg gctgcaggaa
     3781 ttcgatatca agcttatcga taccgtcgac ctcgagtcat gtaattagtt atgtcacgct
     3841 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     3901 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     3961 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     4021 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc
     4081 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     4141 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4201 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4261 cgaatggcgc gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4321 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4381 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4441 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4501 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4561 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4621 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4681 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcct gatgcggtat
     4741 tttctcctta cgcatctgtg cggtatttca caccgcatag gcaagtgcac aaacaatact
     4801 taaataaata ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt
     4861 tgttagagtc ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc
     4921 catttaatct aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat
     4981 gtaagctttc ggggctctct tgccttccaa cccagtcaga aatcgagttc caatccaaaa
     5041 gttcacctgt cccacctgct tctgaatcaa acaagggaat aaacgaatga ggtttctgtg
     5101 aagctgcact gagtagtatg ttgcagtctt ttggaaatac gagtctttta ataactggca
     5161 aaccgaggaa ctcttggtat tcttgccacg actcatctcc atgcagttgg acgatatcaa
     5221 tgccgtaatc attgaccaga gccaaaacat cctccttagg ttgattacga aacacgccaa
     5281 ccaagtattt cggagtgcct gaactatttt tatatgcttt tacaagactt gaaattttcc
     5341 ttgcaataac cgggtcaatt gttctctttc tattgggcac acatataata cccagcaagt
     5401 cagcatcgga atctagagca cattctgcgg cctctgtgct ctgcaagccg caaactttca
     5461 ccaatggacc agaactacct gtgaaattaa taacagacat actccaagct gcctttgtgt
     5521 gcttaatcac gtatactcac gtgctcaata gtcaccaatg ccctccctct tggccctctc
     5581 cttttctttt ttcgaccgaa ttaattctta atcggcaaaa aaagaaaagc tccggatcaa
     5641 gattgtacgt aaggtgacaa gctatttttc aataaagaat atcttccact actgccatct
     5701 ggcgtcataa ctgcaaagta cacatatatt acgatgctgt ctattaaatg cttcctatat
     5761 tatatatata gtaatgtcgt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     5821 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5881 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5941 tttcaccgtc atcaccgaaa cgcgcga
//
