LOCUS       sequence-44436-e        6863 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 6863)
  AUTHORS   Loque D, Lalonde S, Looger LL, von Wiren N, Frommer WB
  TITLE     A cytosolic trans-activation domain essential for ammonium uptake.
  JOURNAL   Nature. 2007 Mar 8;446(7132):195-8. Epub 2007 Feb 11.
  PUBMED    17293878
REFERENCE   2  (bases 1 to 6863)
  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: "Nature."; 
            date: "2007-03-8"; volume: "446(7132)"; pages: "195-8. Epub 2007 Feb
            11"
COMMENT     Sequence Label: pDRf1
FEATURES             Location/Qualifiers
     source          1..6863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..79)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(61..79)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      227..414
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(282..306)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(426..443)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(426..442)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(435..457)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      878..2224
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(2335..3138)
                     /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(3139..3354)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3379..3397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3465..3569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3570..3638,3639..4430)
                     /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(3788..3807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4601..5189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5090..5109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5343..5360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5477..5498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5513..5518,5519..5536,5537..5543)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5551..5567
                     /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     5556..5578
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5575..5591
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5575..5591
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(5621..6076)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5758..5779)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5970..5989
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 actagtgaat tcgcggccgc tgcaggtcga cctcgagggg gggcccggta cccaattcgc
       61 cctatagtga gtcgtattac gcgcggatcc agctttggac ttcttcgcca gaggtttggt
      121 caagtctcca atcaaggttg tcggcttgtc taccttgcca gaaatttacg aaaagatgga
      181 aaagggtcaa atcgttggta gatacgttgt tgacacttct aaataagcga atttcttatg
      241 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
      301 tgactcttag gttttaaaac ganaattctt attcttgagt aactctttcc tgtaggtcag
      361 gttgctttct caggtatagc atgaggtcgc tcttattgac cacacctcta ccggcatgcc
      421 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
      481 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
      541 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
      601 ctccttacgc atctgtgcgg tatttcacac cgcataatcg gatcgtactt gttacccatc
      661 attgaatttt gaacatccga acctgggagt tttccctgaa acagatagta tatttgaacc
      721 tgtataataa tatatagtct agcgctttac ggaagacaat gtatgtattt cggttcctgg
      781 agaaactatt gcatctattg cataggtaat cttgcacgtc gcatccccgg ttcattttct
      841 gcgtttccat cttgcacttc aatagcatat ctttgttaac gaagcatctg tgcttcattt
      901 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
      961 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
     1021 atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     1081 tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat
     1141 acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct
     1201 tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca
     1261 ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa
     1321 aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt
     1381 caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa
     1441 cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta
     1501 ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac
     1561 tctatgaata gttcttacta caattttttt gtctaaagag taatactaga gataaacata
     1621 aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt
     1681 atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga
     1741 agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa
     1801 agtgcgtctt cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag
     1861 ctagagaata ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg agcgcttccg
     1921 aaaatgcaac gcgagctgcg cacatacagc tcactgttca cgtcgcacct atatctgcgt
     1981 gttgcctgta tatatatata catgagaaga acggcatagt gcgtgtttat gcttaaatgc
     2041 gtacttatat gcgtctattt atgtaggatg aaaggtagtc tagtacctcc tgtgatatta
     2101 tcccattcca tgcggggtat cgtatgcttc cttcagcact accctttagc tgttctatat
     2161 gctgccactc ctcaattgga ttagtctcat ccttcaatgc tatcatttcc tttgatattg
     2221 gatcgatccg atgataagct gtcaaacatg agaattgggt aataactgat ataattaaat
     2281 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
     2341 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
     2401 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
     2461 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
     2521 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
     2581 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
     2641 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
     2701 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
     2761 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
     2821 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
     2881 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
     2941 agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa
     3001 ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc
     3061 gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt
     3121 atatgtagct ttcgacatga tttatcttcg tttcctgcat gtttttgttc tgtgcagttg
     3181 ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     3241 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     3301 atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagcta
     3361 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3421 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3481 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3541 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3601 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3661 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3721 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3781 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3841 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3901 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3961 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4021 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4081 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4141 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4201 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4261 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4321 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4381 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4441 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4501 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4561 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4621 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4681 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4741 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4801 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4861 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4921 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4981 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5041 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5101 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5161 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5221 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5281 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5341 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5401 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5461 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5521 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5581 acagctatga ccatgattac gccaagctta ccgcatcagg aaattgtaag cgttaatatt
     5641 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     5701 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     5761 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     5821 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     5881 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     5941 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     6001 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     6061 ccgctacagg gcgcgtccat tcgccaagct tcctgaaacg gagaaacata aacaggcatt
     6121 gctgggatca cccatacatc actctgtttt gcctgacctt ttccggtaat ttgaaaacaa
     6181 acccggtctc gaagcggaga tccggcgata attaccgcag aaataaaccc atacacgaga
     6241 cgtagaacca gccgcacatg gccggagaaa ctcctgcgag aatttcgtaa actcgcgcgc
     6301 attgcatctg tatttcctaa tgcggcactt ccaggcctcg atcgagaccg tttatccatt
     6361 gcttttttgt tgtctttttc cctcgttcac agaaagtctg aagaagctat agtagaacta
     6421 tgagcttttt ttgtttctgt tttccttttt ttttttttta cctctgtgga aattgttact
     6481 ctcacactct ttagttcgtt tgtttgtttt gtttattcca attatgaccg gtgacgaaac
     6541 gtggtcgatg gtgggtaccg cttatgctcc cctccattag tttcgattat ataaaaaggc
     6601 caaatattgt attattttca aatgtcctat cattatcgtc taacatctaa tttctcttaa
     6661 attttttctc tttctttcct ataacaccaa tagtgaaaat ctttttttct tctatatcta
     6721 caaaaacttt ttttttctat caacctcgtt gataaatttt ttctttaaca atcgttaata
     6781 attaattaat tggaaaataa ccattttttc tctcttttat acacacattc aaaagaaaga
     6841 aaaaaaatat accccagcct cga
//
