LOCUS       sequence-7822-ex        7382 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 7382)
  AUTHORS   Alberti S, Gitler AD, Lindquist S
  TITLE     A suite of Gateway cloning vectors for high-throughput genetic 
            analysis in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2007 Oct . 24(10):913-9.
  PUBMED    17583893
REFERENCE   2  (bases 1 to 7382)
  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: "Yeast. 2007
            Oct . 24(10):913-9."
COMMENT     Sequence Label: pAG304GAL-ccdB-ECFP
FEATURES             Location/Qualifiers
     source          1..7382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        185..465
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             466..1140
                     /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"
     rep_origin      complement(1237..1692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1374..1395)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1586..1605
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1822..1844
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1836..1853
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1837..1853
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1860..1879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1860..1878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1898..2339
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1898..2015
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2310..2333
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     2354..2370
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2354..2370
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2378..2502
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2527..2532,2533..2550,2551..2557)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2611..3270
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2677..2696)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3615..3920
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3799..3818
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3961..4085)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(4117..4119,4120..4122,4123..4833)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4162..4183)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4423..4442)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4770..4791
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      4852..5099
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4868..4886)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(5122..5140)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5161..5177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5161..5177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5174..5196)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5185..5201
                     /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(5209..5215,5216..5233,5234..5239))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5392..5409)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5563..6151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6322..7113,7114..7182))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6945..6964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7183..7287)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7355..7373
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accaaacgac attactatat atataatata ggaagcattt aatagacagc atcgtaatat
      241 atgtgtactt tgcagttatg acgccagatg gcagtagtgg aagatattct ttattgaaaa
      301 atagcttgtc accttacgta caatcttgat ccggagcttt tctttttttg ccgattaaga
      361 attaattcgg tcgaaaaaag aaaaggagag ggccaagagg gagggcattg gtgactattg
      421 agcacgtgag tatacgtgat taagcacaca aaggcagctt ggagtatgtc tgttattaat
      481 ttcacaggta gttctggtcc attggtgaaa gtttgcggct tgcagagcac agaggccgca
      541 gaatgtgctc tagattccga tgctgacttg ctgggtatta tatgtgtgcc caatagaaag
      601 agaacaattg acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat
      661 agttcaggca ctccgaaata cttggttggc gtgtttcgta atcaacctaa ggaggatgtt
      721 ttggctctgg tcaatgatta cggcattgat atcgtccaac tgcatggaga tgagtcgtgg
      781 caagaatacc aagagttcct cggtttgcca gttattaaaa gactcgtatt tccaaaagac
      841 tgcaacatac tactcagtgc agcttcacag aaacctcatt cgtttattcc cttgtttgat
      901 tcagaagcag gtgggacagg tgaacttttg gattggaact cgatttctga ctgggttgga
      961 aggcaagaga gccccgaaag cttacatttt atgttagctg gtggactgac gccagaaaat
     1021 gttggtgatg cgcttagatt aaatggcgtt attggtgttg atgtaagcgg aggtgtggag
     1081 acaaatggtg taaaagactc taacaaaata gcaaatttcg tcaaaaatgc taagaaatag
     1141 gttattactg agtagtattt atttaagtat tgtttgtgca cttgcctgcg gtgtgaaata
     1201 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaaacgtt aatattttgt
     1261 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1321 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1381 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1441 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1501 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1561 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1621 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1681 tacagggcgc gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1741 tgcgggcctc ttcgctatta cgccagctgg cgaagggggg atgtgctgca aggcgattaa
     1801 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt
     1861 aatacgactc actatagggc gaattggagc tctagtacgg attagaagcc gccgagcggg
     1921 cgacagccct ccgacggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc
     1981 tgaaacgcag atgtgcctcg cgccgcactg ctccgaacaa taaagattct acaatactag
     2041 cttttatggt tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatta
     2101 acgaatcaaa ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg
     2161 ggtaattaat cagcgaagcg atgatttttg atctattaac agatatataa atggaaaagc
     2221 tgcataacca ctttaactaa tactttcaac attttcagtt tgtattactt cttattcaaa
     2281 tgtcataaaa gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga
     2341 aaaaaccccg gattctagaa ctagtggatc ccccatcaca agtttgtaca aaaaagctga
     2401 acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca
     2461 gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc
     2521 ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg
     2581 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     2641 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     2701 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     2761 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     2821 ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     2881 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     2941 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     3001 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     3061 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     3121 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     3181 gttcatcatg ccgtctgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     3241 tactgcgatg agtggcaggg cggggcgtaa acgccgcgtg gatccggctt actaaaagcc
     3301 agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg
     3361 tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg
     3421 acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag
     3481 cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca
     3541 ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa
     3601 caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc
     3661 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc
     3721 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata
     3781 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
     3841 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
     3901 tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac
     3961 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
     4021 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa
     4081 gtggtgatgg gctgcaggaa ttcgatatca agcttaatgg tgagcaaggg cgaggagctg
     4141 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     4201 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     4261 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctggggc
     4321 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     4381 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     4441 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     4501 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacatcagc
     4561 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     4621 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     4681 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     4741 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     4801 gggatcactc tcggcatgga cgagctgtac aagctgtaca agtaactcga gtcatgtaat
     4861 tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag
     4921 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     4981 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     5041 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg
     5101 gccggtaccc agcttttgtt ccctttagtg agggttaatt ccgagcttgg cgtaatcatg
     5161 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acataggagc
     5221 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg aggtaactca cattaattgc
     5281 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5341 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5401 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5461 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5521 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctcggccc
     5581 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5641 ataaagatac caggcgttcc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5701 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     5761 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5821 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5881 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5941 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6001 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6061 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6121 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6181 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6241 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6301 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6361 ctgtctattt cgttcatcca tagttgcctg actgcccgtc gtgtagataa ctacgatacg
     6421 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6481 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6541 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6601 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6661 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6721 ccccatgttg tgaaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6781 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6841 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6901 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6961 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7021 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7081 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7141 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7201 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7261 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     7321 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     7381 tc
//
