LOCUS       sequence-122735-        8450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TCP4 expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8450)
  AUTHORS   Rubio-Somoza I, Weigel D
  TITLE     Coordination of flower maturation by a regulatory circuit of three 
            microRNAs.
  JOURNAL   PLoS Genet. 2013 Mar;9(3):e1003374. doi: 
            10.1371/journal.pgen.1003374. Epub 2013 Mar 28.
  PUBMED    23555288
REFERENCE   2  (bases 1 to 8450)
  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: "PLoS 
            Genet."; date: "2013-03"; volume: "9(3):e1003374. doi"; pages: " 
            10.1371/journal.pgen.1003374. Epub 2013 Mar 28"
COMMENT     Sequence Label: pIR190
FEATURES             Location/Qualifiers
     source          1..8450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    81..102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(117..122,123..140,141..147)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    155..171
                     /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     160..182
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     179..195
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     179..195
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        216..234
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1018..1722
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1734..1754
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1764..2105
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     protein_bind    2121..2145
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3461..3485)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      3685..3872
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3740..3764)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4024..4043)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4025..4043)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4050..4067)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4050..4066)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4059..4081)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4207..4662
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4294..4313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4504..4525
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4765..5439)
                     /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"
     promoter        complement(5440..5720)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5741..5760)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5860..5882
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5920..5938)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5979..6482
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6519..6623
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6624..6692,6693..7484)
                     /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(6842..6861)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7655..8243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8144..8163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8397..8414
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
       61 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      121 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      181 ggaaacagct atgaccatga ttacgccaag ctcgaaatta accctcacta aagggaacaa
      241 aagctggtac cgggcccccc ctcgagatcc gggatcgaag aaatgatggt aaatgaaata
      301 ggaaatcaag gagcatgaag gcaaaagaca aatataaggg tcgaacgaaa aataaagtga
      361 aaagtgttga tatgatgtat ttggctttgc ggcgccgaaa aaacgagttt acgcaattgc
      421 acaatcatgc tgactctgtg gcggacccgc gctcttgccg gcccggcgat aacgctgggc
      481 gtgaggctgt gcccggcgga gttttttgcg cctgcatttt ccaaggttta ccctgcgcta
      541 aggggcgaga ttggagaagc aataagaatg ccggttgggg ttgcgatgat gacgaccacg
      601 acaactggtg tcattattta agttgccgaa agaacctgag tgcatttgca acatgagtat
      661 actagaagaa tgagccaaga cttgcgagac gcgagtttgc cggtggtgcg aacaatagag
      721 cgaccatgac cttgaaggtg agacgcgcat aaccgctaga gtactttgaa gaggaaacag
      781 caatagggtt gctaccagta taaatagaca ggtacataca acactggaaa tggttgtctg
      841 tttgagtacg ctttcaattc atttgggtgt gcactttatt atgttacaat atggaaggga
      901 actttacact tctcctatgc acatatatta attaaagtcc aatgctagta gagaaggggg
      961 gtaacacccc tccgcgctct tttccgattt ttttctaaac cgtggaatat ttcggatatc
     1021 cttttgttgt ttccgggtgt acaatatgga cttcctcttt tctggcaacc aaacccatac
     1081 atcgggattc ctataatacc ttcgttggtc tccctaacat gtaggtggcg gaggggagat
     1141 atacaataga acagatacca gacaagacat aatgggctaa acaagactac accaattaca
     1201 ctgcctcatt gatggtggta cataacgaac taatactgta gccctagact tgatagccat
     1261 catcatatcg aagtttcact accctttttc catttgccat ctattgaagt aataataggc
     1321 gcatgcaact tcttttcttt ttttttcttt tctctctccc ccgttgttgt ctcaccatat
     1381 ccgcaatgac aaaaaaatga tggaagacac taaaggaaaa aattaacgac aaagacagca
     1441 ccaacagatg tcgttgttcc agagctgatg aggggtatct cgaagcacac gaaacttttt
     1501 ccttccttca ttcacgcaca ctactctcta atgagcaacg gtatacggcc ttccttccag
     1561 ttacttgaat ttgaaataaa aaaaagtttg ctgtcttgct atcaagtata aatagacctg
     1621 caattattaa tcttttgttt cctcgtcatt gttctcgttc cctttcttcc ttgtttcttt
     1681 ttctgcacaa tatttcaagc tataccaagc atacaatcaa ctccaagctt atgcccaaga
     1741 agaagcggaa ggtctcgagc ggcgccaatt ttaatcaaag tgggaatatt gctgatagct
     1801 cattgtcctt cactttcact aacagtagca acggtccgaa cctcataaca actcaaacaa
     1861 attctcaagc gctttcacaa ccaattgcct cctctaacgt tcatgataac ttcatgaata
     1921 atgaaatcac ggctagtaaa attgatgatg gtaataattc aaaaccactg tcacctggtt
     1981 ggacggacca aactgcgtat aacgcgtttg gaatcactac agggatgttt aataccacta
     2041 caatggatga tgtatataac tatctattcg atgatgaaga taccccacca aacccaaaaa
     2101 aagagggtgg gtcgaatcaa acaagtttgt acaaaaaagc aggctccgaa ttcgcccttc
     2161 ccggatccat gtctgacgac caattccatc acccgccgcc tccttcttca atgaggcacc
     2221 gttctacgtc ggatgcggcg gacggcggct gcggcgagat tgttgaggtg caaggtggtc
     2281 acattgttcg gtctaccgga agaaaagacc gccacagcaa agtctgcacg gctaaagggc
     2341 cacgtgaccg gcgcgtgaga ctctctgctc acacggcgat tcagttttac gatgttcaag
     2401 acaggcttgg tttcgaccga cctagcaaag ccgttgattg gcttatcaaa aaggctaaga
     2461 cttccattga cgagctcgct gagcttcctc cctggaatcc cgccgatgca attcgcctag
     2521 ccgctgctaa cgctaaaccc agaagaacca ccgccaaaac ccaaatctct ccgtctccgc
     2581 caccgccgca acagcaacaa caacaacaac agcttcagtt cggtgttggc ttcaacggag
     2641 gaggagcaga gcatccgagt aacaacgagt cgagttttct cccgccgtca atggattcag
     2701 attcgatagc tgacactata aagtcgtttt ttccggtgat tggctcttca acggaggctc
     2761 cttcgaatca taaccttatg cacaactatc atcatcagca tccgccggat ttgctttctc
     2821 gaactaatag ccaaaaccaa gatctccgtc tctcgctgca atcgttcccg gatggtccac
     2881 cgtcgcttct gcaccaccaa catcaccacc acacctctgc ttccgcctcc gagcctactc
     2941 tgttctacgg acagagcaat ccgttagggt ttgacacatc gagttgggag cagcagtcgt
     3001 cggaattcgg aaggattcag agactagtgg cttggaacag cggcggtggc ggcggagcaa
     3061 ccgatacagg aaacggagga gggtttctgt tcgctcctcc tactccttca acgacgtcgt
     3121 ttcagccagt tcttggccaa agccaacagc tttattctca gaggggtccc cttcagtcca
     3181 gttacagtcc catgatccgt gcttggtttg atcctcacca tcatcaccaa tccatctcca
     3241 ccgacgatct caaccaccac catcaccttc ctccaccggt tcaccaatca gcaatccccg
     3301 gaatcggatt cgcctcaggt gaattctctt cgggttttcg cataccagca cggtttcagg
     3361 gccaagaaga ggagcagcac gacggtctca ctcacaagcc gtcctctgct tcctctattt
     3421 ctcgccattg aaaaggccct gtcgacgaag ggcgaattcg acccagcttt cttgtacaaa
     3481 gtggtttgat ggccgctaag taagtaagac gtcgagctct aagtaagtaa cggccgccac
     3541 cgcggtggag ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc
     3601 ggcttgtcta ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga
     3661 tacgttgttg acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     3721 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     3781 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat
     3841 gaggtcgctc ttattgacca cacctctacc ggcatgccga gcaaatgcct gcaaatcgct
     3901 ccccatttca cccaattgta gatatgctaa ctccagcaat gagttgatga atctcggtgt
     3961 gtattttatg tcctcagagg acaacacctg ttgtaatcgt tcttccacac ggatcccaat
     4021 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     4081 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4141 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4201 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4261 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4321 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4381 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4441 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4501 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4561 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4621 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     4681 ccttacgcat ctgtgcggta tttcacaccg catagatcgg caagtgcaca aacaatactt
     4741 aaataaatac tactcagtaa taacctattt cttagcattt ttgacgaaat ttgctatttt
     4801 gttagagtct tttacaccat ttgtctccac acctccgctt acatcaacac caataacgcc
     4861 atttaatcta agcgcatcac caacattttc tggcgtcagt ccaccagcta acataaaatg
     4921 taagctttcg gggctctctt gccttccaac ccagtcagaa atcgagttcc aatccaaaag
     4981 ttcacctgtc ccacctgctt ctgaatcaaa caagggaata aacgaatgag gtttctgtga
     5041 agctgcactg agtagtatgt tgcagtcttt tggaaatacg agtcttttaa taactggcaa
     5101 accgaggaac tcttggtatt cttgccacga ctcatctcca tgcagttgga cgatatcaat
     5161 gccgtaatca ttgaccagag ccaaaacatc ctccttaggt tgattacgaa acacgccaac
     5221 caagtatttc ggagtgcctg aactattttt atatgctttt acaagacttg aaattttcct
     5281 tgcaataacc gggtcaattg ttctctttct attgggcaca catataatac ccagcaagtc
     5341 agcatcggaa tctagagcac attctgcggc ctctgtgctc tgcaagccgc aaactttcac
     5401 caatggacca gaactacctg tgaaattaat aacagacata ctccaagctg cctttgtgtg
     5461 cttaatcacg tatactcacg tgctcaatag tcaccaatgc cctccctctt ggccctctcc
     5521 ttttcttttt tcgaccgaat taattcttaa tcggcaaaaa aagaaaagct ccggatcaag
     5581 attgtacgta aggtgacaag ctatttttca ataaagaata tcttccacta ctgccatctg
     5641 gcgtcataac tgcaaagtac acatatatta cgatgctgtt ctattaaatg cttcctatat
     5701 tatatatata gtaatgtcgt gatctatggt gcactctcag tacaatctgc tctgatgccg
     5761 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5821 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5881 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5941 tataggttaa tgtcatgata ataatggttt cttagacgga tcgcttgcct gtaacttaca
     6001 cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt
     6061 ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga
     6121 agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata
     6181 tttattagac aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg
     6241 taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat
     6301 taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga
     6361 gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct
     6421 atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg
     6481 atgaaaagga cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6541 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6601 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6661 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6721 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6781 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6841 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6901 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6961 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7021 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7081 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7141 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7201 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7261 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7321 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7381 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7441 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7501 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7561 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7621 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7681 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7741 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7801 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7861 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7921 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7981 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8041 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8101 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8161 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8221 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8281 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8341 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8401 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
//
