LOCUS       sequence-104872-        5364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS403 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 5364)
  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 5364)
  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: pAV103
FEATURES             Location/Qualifiers
     source          1..5364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..22
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     122..141
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        288..474
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             475..1134
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1394..1849)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1531..1552)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1743..1762
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1979..2001
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1993..2010
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1994..2010
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2020..2039
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2020..2038
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2063..2079
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2064..2080
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2164..2841)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2848..2859
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2877..2893
                     /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)."
     promoter        complement(join(2901..2907,2908..2925,2926..2931))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2946..2967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3082..3098)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3082..3098)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3135..3155)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3135..3153)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3174..3190)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3174..3190)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3198..3214
                     /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)."
     promoter        complement(join(3222..3228,3229..3246,3247..3252))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3405..3422)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3576..4164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3656..3675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4335..5126,5127..5195))
                     /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     4958..4977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5196..5300)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataggc gtatcacgag gccctttgtc acagcttgtc tgtaagcgga tgccgggagc
       61 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
      121 gcggcatcag agcagattgt actgagagtg caccataaat tcccgtttta agagcttggt
      181 gagcgctagg agtcactgcc aggtatcgtt tgaacacggc attagtcagg gaagtcataa
      241 cacagtcctt tcccgcaatt ttctttttct attactcttg gcctcctcta gtacactcta
      301 tattttttta tgcctcggta atgattttca tttttttttt tcccctagcg gatgactctt
      361 tttttttctt agcgattggc attatcacat aatgaattat acattatata aagtaatgtg
      421 atttcttcga agaatatact aaaaaatgag caggcaagat aaacgaaggc aaagatgaca
      481 gagcagaaag ccctagtaaa gcgtattaca aatgaaacca agattcagat tgcgatctct
      541 ttaaagggtg gtcccctagc gatagagcac tcgatcttcc cagaaaaaga ggcagaagca
      601 gtagcagaac aggccacaca atcgcaagtg attaacgtcc acacaggtat agggtttctg
      661 gaccatatga tacatgctct ggccaagcat tccggctggt cgctaatcgt tgagtgcatt
      721 ggtgacttac acatagacga ccatcacacc actgaagact gcgggattgc tctcggtcaa
      781 gcttttaaag aggccctact ggcgcgtgga gtaaaaaggt ttggatcagg atttgcgcct
      841 ttggatgagg cactttccag agcggtggta gatctttcga acaggccgta cgcagttgtc
      901 gaacttggtt tgcaaaggga gaaagtagga gatctctctt gcgagatgat cccgcatttt
      961 cttgaaagct ttgcagaggc tagcagaatt accctccacg ttgattgtct gcgaggcaag
     1021 aatgatcatc accgtagtga gagtgcgttc aaggctcttg cggttgccat aagagaagcc
     1081 acctcgccca atggtaccaa cgatgttccc tccaccaaag gtgttcttat gtagtgacac
     1141 cgattattta aagctgcagc atacgatata tatacatgtg tatatatgta tacctatgaa
     1201 tgtcagtaag tatgtatacg aacagtatga tactgaagat gacaaggtaa tgcatcattc
     1261 tatacgtgtc attctgaacg aggcgcgctt tccttttttc tttttgcttt ttcttttttt
     1321 ttctcttgaa ctcgacggat ctatgcggtg tgaaataccg cacagatgcg taaggagaaa
     1381 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     1441 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     1501 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     1561 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     1621 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     1681 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     1741 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     1801 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
     1861 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     1921 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     1981 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
     2041 gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc
     2101 ctgcagcccc cagtagagac cgcctggctc tagtagcgat ctacactagc actatcagcg
     2161 ttattaagca ccggtggagt gacgaccttc agcacgttcg tactgttcaa cgatggtgta
     2221 gtcttcgttg tgggaggtga tgtccagttt gatgtcggtt ttgtaagcac ccggcagctg
     2281 aaccggtttt ttagccatgt aggtggtttt aacttcagcg tcgtagtgac caccgtcttt
     2341 cagtttcaga cgcattttga tttcaccttt cagagcaccg tcttccgggt acatacgttc
     2401 ggtggaagct tcccaaccca tggttttttt ctgcataacc ggaccgtcgg acgggaagtt
     2461 ggtaccacgc agtttaactt tgtagatgaa ctcaccgtct tgcagggagg agtcctgggt
     2521 aacggtaaca acaccaccgt cttcgaagtt cataacacgt tcccatttga aaccttccgg
     2581 gaaggacagt ttcaggtagt ccgggatgtc agccgggtgt ttaacgtaag ctttggaacc
     2641 gtactggaac tgcggggaca ggatgtccca agcgaacggc agcggaccac ctttggtaac
     2701 tttcagttta gcggtctggg taccttcgta cggacgacct tcaccttcac cttcgatttc
     2761 gaactcgtga ccgttaacgg aaccttccat acgaactttg aaacgcatga actctttgat
     2821 aacgtcttcg gaggaagcca tctagtattt ctcctctttc tctagtatgt gtgaaattgt
     2881 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2941 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3001 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3061 cgtattgggt ctcattttgg ggatccacta gttctagagc ggccgccacc gcggtggagc
     3121 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     3181 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc
     3241 ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc
     3301 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3361 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3421 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3481 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3541 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3601 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3661 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3721 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3781 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3841 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3901 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3961 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4021 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4081 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4141 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4201 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4261 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4321 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4381 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4441 ttaccatctg gccccagtgc tgcaatgata ccgcgagatc cacgctcacc ggctccagat
     4501 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4561 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4621 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4681 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4741 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4801 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4861 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4921 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4981 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5041 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5101 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5161 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5221 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5281 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     5341 attattatca tgacattaac ctat
//
