LOCUS       sequence-44435-e        6406 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 6406)
  AUTHORS   Meyer A, Eskandari S, Grallath S, Rentsch D
  TITLE     AtGAT1, a high affinity transporter for gamma-aminobutyric acid in 
            Arabidopsis thaliana.
  JOURNAL   J Biol Chem. 2006 Mar 17;281(11):7197-204. Epub 2006 Jan 10.
  PUBMED    16407306
REFERENCE   2  (bases 1 to 6406)
  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: "J Biol 
            Chem."; date: "2006-03-17"; volume: "281(11)"; pages: "7197-204. 
            Epub 2006 Jan 10"
COMMENT     Sequence Label: pDR196
FEATURES             Location/Qualifiers
     source          1..6406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..60)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(45..61)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(85..104)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(86..104)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      252..439
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(307..331)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(451..468)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(451..467)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(460..482)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      903..2249
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(2360..3163)
                     /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(3164..3379)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3404..3422)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3490..3594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3595..3663,3664..4455)
                     /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(3813..3832)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4626..5214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5115..5134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5368..5385
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5502..5523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5538..5543,5544..5561,5562..5568)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5576..5592
                     /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     5581..5603
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5600..5616
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5600..5616
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctagtggatc ccccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg
       61 agggggggcc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg gatccagctt
      121 tggacttctt cgccagaggt ttggtcaagt ctccaatcaa ggttgtcggc ttgtctacct
      181 tgccagaaat ttacgaaaag atggaaaagg gtcaaatcgt tggtagatac gttgttgaca
      241 cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa
      301 aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaraa ttcttattct
      361 tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagcatgag gtcgctctta
      421 ttgaccacac ctctaccggc atgccaattc actggccgtc gttttacaac gtcgtgactg
      481 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
      541 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
      601 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
      661 aatcggatcg tacttgttac ccatcattga attttgaaca tccgaacctg ggagttttcc
      721 ctgaaacaga tagtatattt gaacctgtat aataatatat agtctagcgc tttacggaag
      781 acaatgtatg tatttcggtt cctggagaaa ctattgcatc tattgcatag gtaatcttgc
      841 acgtcgcatc cccggttcat tttctgcgtt tccatcttgc acttcaatag catatctttg
      901 ttaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt
      961 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat
     1021 tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct
     1081 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc
     1141 gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga
     1201 gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa
     1261 tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt
     1321 ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac
     1381 tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg
     1441 atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc
     1501 agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta
     1561 cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta
     1621 aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa
     1681 ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga
     1741 gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag
     1801 tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc
     1861 gctctgaagt tcctatactt tctagctaga gaataggaac ttcggaatag gaacttcaaa
     1921 gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact
     1981 gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc
     2041 atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg
     2101 tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca
     2161 gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc
     2221 aatgctatca tttcctttga tattggatcg atccgatgat aagctgtcaa acatgagaat
     2281 tgggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt atacatgcat
     2341 ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat gcttcccagc
     2401 ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca aatagtcctc
     2461 ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt ctatactgtt
     2521 gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc aaccaatcgt
     2581 aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca aaatctttgt
     2641 cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg gagcccttgc
     2701 atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct tctgccgcct
     2761 gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta atgtctgccc
     2821 attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta ccaatgtcag
     2881 caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc tttagcggct
     2941 taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt agtaaacaaa
     3001 ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga acatccaatg
     3061 aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca gcaacaggac
     3121 taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat cttcgtttcc
     3181 tgcatgtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat gtttcttcaa
     3241 cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct
     3301 gttcggagat taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa aaagaataaa
     3361 aaaaaaatga tgaattgaaa agctaattct tgaagacgaa agggcctcgt gatacgccta
     3421 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3481 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3541 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3601 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3661 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3721 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3781 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3841 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3901 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3961 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4021 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4081 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4141 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4201 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4261 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4321 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4381 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4441 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4501 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4741 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4861 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4921 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4981 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5041 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5101 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5161 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5221 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5281 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5341 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5401 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     5461 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     5521 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     5581 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttcctgaa
     5641 acggagaaac ataaacaggc attgctggga tcacccatac atcactctgt tttgcctgac
     5701 cttttccggt aatttgaaaa caaacccggt ctcgaagcgg agatccggcg ataattaccg
     5761 cagaaataaa cccatacacg agacgtagaa ccagccgcac atggccggag aaactcctgc
     5821 gagaatttcg taaactcgcg cgcattgcat ctgtatttcc taatgcggca cttccaggcc
     5881 tcgatcgaga ccgtttatcc attgcttttt tgttgtcttt ttccctcgtt cacagaaagt
     5941 ctgaagaagc tatagtagaa ctatgagctt tttttgtttc tgttttcctt tttttttttt
     6001 ttacctctgt ggaaattgtt actctcacac tctttagttc gtttgtttgt tttgtttatt
     6061 ccaattatga ccggtgacga aacgtggtcg atggtgggta ccgcttatgc tcccctccat
     6121 tagtttcgat tatataaaaa ggccaaatat tgtattattt tcaaatgtcc tatcattatc
     6181 gtctaacatc taatttctct taaatttttt ctctttcttt cctataacac caatagtgaa
     6241 aatctttttt tcttctatat ctacaaaaac tttttttttc tatcaacctc gttgataaat
     6301 tttttcttta acaatcgtta ataattaatt aattggaaaa taaccatttt ttctctcttt
     6361 tatacacaca ttcaaaagaa agaaaaaaaa tataccccag cctcga
//
