LOCUS       sequence-331731-        6704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS423 backbone, contains an RFP flanked by BsaI sites with CAGT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6704)
  AUTHORS   Agmon N, Mitchell LA, Cai Y, Ikushima S, Chuang J, Zheng A, Choi WJ,
            Martin JA, Caravelli K, Stracquadanio G, Boeke JD
  TITLE     Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae 
            Transcription Units.
  JOURNAL   ACS Synth Biol. 2015 Jul 17;4(7):853-9. doi: 10.1021/sb500372z. Epub
            2015 Mar 23.
  PUBMED    25756291
REFERENCE   2  (bases 1 to 6704)
  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"; doi: "10.1021/sb500372z";
            journalName: "ACS Synth Biol"; date: "2015-07-17- 17"; volume: "4"; 
            issue: "7"; pages: "853-9"
COMMENT     Sequence Label: pAV123
FEATURES             Location/Qualifiers
     source          1..6704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     36..56
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        38..56
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     93..109
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     93..109
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    224..245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(260..265,266..283,284..290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    298..314
                     /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)."
     RBS             332..343
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             350..1027
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1111..1127)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1112..1128)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1152..1171)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1153..1171)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1181..1198)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1181..1197)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1190..1212)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1336..1791
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1423..1442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1633..1654
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2051..2713)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(2714..2901)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(3048..3067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3167..3185)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      3226..4568
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    3558..3605
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        4595..4699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4700..4768,4769..5560)
                     /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(4918..4937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5731..6319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6220..6239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6473..6490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6607..6628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6643..6648,6649..6666,6667..6673)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6681..6697
                     /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)."
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
       61 aaaagctgga gctccaccgc ggtggcggcc gctctagaac tagtggatcc ccaaaatgag
      121 acccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
      181 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      241 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      301 tgagcggata acaatttcac acatactaga gaaagaggag aaatactaga tggcttcctc
      361 cgaagacgtt atcaaagagt tcatgcgttt caaagttcgt atggaaggtt ccgttaacgg
      421 tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta cccagaccgc
      481 taaactgaaa gttaccaaag gtggtccgct gccgttcgct tgggacatcc tgtccccgca
      541 gttccagtac ggttccaaag cttacgttaa acacccggct gacatcccgg actacctgaa
      601 actgtccttc ccggaaggtt tcaaatggga acgtgttatg aacttcgaag acggtggtgt
      661 tgttaccgtt acccaggact cctccctgca agacggtgag ttcatctaca aagttaaact
      721 gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca tgggttggga
      781 agcttccacc gaacgtatgt acccggaaga cggtgctctg aaaggtgaaa tcaaaatgcg
      841 tctgaaactg aaagacggtg gtcactacga cgctgaagtt aaaaccacct acatggctaa
      901 aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc aaactggaca tcacctccca
      961 caacgaagac tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc actccaccgg
     1021 tgcttaataa cgctgatagt gctagtgtag atcgctacta gagccaggcg gtctctactg
     1081 ggggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagg gggggcccgg
     1141 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     1201 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1261 tcgccagctg gcgtaatagc gaagaggccc gcaccgcgcc cttcccaaca gttgcgcagc
     1321 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1381 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1441 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1501 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1561 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1621 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1681 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1741 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     1801 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagatccgt cgagttcaag
     1861 agaaaaaaaa agaaaaagca aaaagaaaaa aggaaagcgc gcctcgttca gaatgacacg
     1921 tatagaatga tgcattacct tgtcatcttc agtatcatac tgttcgtata catacttact
     1981 gacattcata ggtatacata tatacacatg tatatatatc gtatgctgca gctttaaata
     2041 atcggtgtca ctacataaga acacctttgg tggagggaac atcgttggta ccattgggcg
     2101 aggtggcttc tcttatggca accgcaagag ccttgaacgc actctcacta cggtgatgat
     2161 cattcttgcc tcgcagacaa tcaacgtgga gggtaattct gctagcctct gcaaagcttt
     2221 caagaaaatg cgggatcatc tcgcaagaga gatctcctac tttctccctt tgcaaaccaa
     2281 gttcgacaac tgcgtacggc ctgttcgaaa gatctaccac cgctctggaa agtgcctcat
     2341 ccaaaggcgc aaatcctgat ccaaaccttt ttactccacg cacggcccct agggcctctt
     2401 taaaagcttg accgagagca atcccgcagt cttcagtggt gtgatggtcg tctatgtgta
     2461 agtcaccaat gcactcaacg attagcgacc agccggaatg cttggccaga gcatgtatca
     2521 tatggtccag aaaccctata cctgtgtgga cgttaatcac ttgcgattgt gtggcctgtt
     2581 ctgctactgc ttctgcctct ttttctggga agatcgagtg ctctatcgct aggggaccac
     2641 cctttaaaga gatcgcaatc tgaatcttgg tttcatttgt aatacgcttt actagggctt
     2701 tctgctctgt catctttgcc ttcgtttatc ttgcctgctc attttttagt atattcttcg
     2761 aagaaatcac attactttat ataatgtata attcattatg tgataatgcc aatcgctaag
     2821 aaaaaaaaag agtcatccgc taggtggaaa aaaaaaaatg aaaatcatta ccgaggcata
     2881 aaaaaatata gagtgtacta gaggaggcca agagtaatag aaaaagaaaa ttgcgggaaa
     2941 ggactgtgtt atgacttccc tgactaatgc cgtgttcaaa cgatacctgg cagtgactcc
     3001 tagcgctcac caagctctta aaacgggaat ttatggtgca ctctcagtac aatctgctct
     3061 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3121 gcttgtctgc tcccggcatc cgcttacaga caagctgtga caaagggcct cgtgatacgc
     3181 ctatttttat aggttaatgt catgataata atggtttctt agtatgatcc aatatcaaag
     3241 gaaatgatag cattgaagga tgagactaat ccaattgagg agtggcagca tatagaacag
     3301 ctaaagggta gtgctgaagg aagcatacga taccccgcat ggaatgggat aatatcacag
     3361 gaggtactag actacctttc atcctacata aatagacgca tataagtacg catttaagca
     3421 taaacacgca ctatgccgtt cttctcatgt atatatatat acaggcaaca cgcagatata
     3481 ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg ttgcattttc ggaagcgctc
     3541 gttttcggaa acgctttgaa gttcctattc cgaagttcct attctctaga aagtatagga
     3601 acttcagagc gcttttgaaa accaaaagcg ctctgaagac gcactttcaa aaaaccaaaa
     3661 acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac aaacattgct
     3721 caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct acccatccac
     3781 ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt ttatctctag
     3841 tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa tcgaaaacaa
     3901 tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa gaaaccgttc
     3961 ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac aaagtatgcg
     4021 caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa atgcacccgc
     4081 agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta tggaagagaa
     4141 aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa aaagttatca
     4201 agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg ctttgttaga
     4261 aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga ttctttgttg
     4321 gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt
     4381 ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc acagattctt
     4441 cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca gctcagattc
     4501 tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg aagcacagat
     4561 gcttcgttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4621 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4681 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4741 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4801 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4861 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4921 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4981 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5041 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5101 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5161 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5221 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5281 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5341 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5401 agccggtgag cgtggatctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5461 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5521 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5581 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5641 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5701 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5761 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5821 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5881 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5941 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6001 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6061 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6481 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6541 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6601 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6661 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt caca
//
