LOCUS       Exported                9014 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Express SSN6.
ACCESSION   .
VERSION     .
KEYWORDS    pSSN6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9014)
  AUTHORS   Xia Z, Turner GC, Hwang CS, Byrd C, Varshavsky A
  TITLE     Amino acids induce peptide uptake via accelerated degradation of 
            CUP9, the transcriptional repressor of the PTR2 peptide transporter.
  JOURNAL   J Biol Chem. 2008 Oct 24;283(43):28958-68. doi: 
            10.1074/jbc.M803980200. Epub 2008 Aug 15.
  PUBMED    18708352
REFERENCE   2  (bases 1 to 9014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9014
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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        191..471
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             472..1146
                     /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(1249..1704)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1386..1407)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1598..1617
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1830..1852
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1844..1861
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1845..1861
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1868..1887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1868..1886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1919..1935
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1919..1935
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6196..6212)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6197..6213)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(6241..6259)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6280..6296)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6280..6296)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6293..6315)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6304..6320
                     /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(6328..6358)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6373..6394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6511..6528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6682..7270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6762..6781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7441..8301)
                     /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     8064..8083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8302..8406)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8443..8946
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8987..9005
                     /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 accatagatc acgacattac tatatatata atataggaag catttaatag aacagcatcg
      241 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat
      301 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga
      361 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
      421 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
      481 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
      541 gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat
      601 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
      661 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
      721 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
      781 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
      841 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
      901 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg
      961 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca
     1021 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt
     1081 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag
     1141 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg
     1201 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
     1261 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1321 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1381 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1441 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1501 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1561 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1621 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1681 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg
     1741 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     1801 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     1861 tgaattgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc
     1921 tagaactagt ggatcccgaa gataaaaact aaagcacatc cgatctgagt tttgagctgg
     1981 gcggcaaaaa caaaactgtc agtggctggc tgatacaggg cacgtgagag gtcactgagg
     2041 acgaagggtg cggctcaccc ggattggctg agtcaccttc atcgcgtcca cgtcactcct
     2101 cgttcgcttt ttttttccct tttcttcttt tgtgcaccac tcagggcacg agaaaagctt
     2161 atttttttcg tgcacactcc gttcttagtg gttgttccac atcttttttg tgcttgtgcg
     2221 tttgccaatg ctaaaatgga aattgcctta taacgtgggc catgtcatgc cgtgtcctct
     2281 cgtttcgtaa aacttaccac acgtaccgga ctgttccgtc ctgtctcgta tcgcacgggt
     2341 gccgtatcgg ctctaattat tttatctctc tattttcttt cttttctctg cgctactcct
     2401 ttctcgatcg ttgctactcc cgtcgctagc cactggtctc ccgcgtactg tactccatct
     2461 ttttttggcg tttttcccct atccaactcg aacaaggttt gtttaaattt attttatttt
     2521 tcttttcttc ggtcggtcgt tcttttccct tccgattatc aaagcaaaag cgcatttttt
     2581 tctttgtctt tttgtttttt gtttcctgtt ctctgttttt ttacaaacca cgtcaggagt
     2641 tcaattgaga gaactagaat caacaaagcc aaatacgaca acgtcactag tctttgaacc
     2701 agaggcgtat tcccgttacc tcttttccca tatttctgtt tttctttttc actgctataa
     2761 gcctttagac tagtactaca actacaacag caacaacaac aaacaaaaca cgactggaaa
     2821 aaaaaaatta ggaaaaatga atccgggcgg tgaacaaaca ataatggaac aacccgctca
     2881 acagcaacaa caacagcaac aacaacagca gcaacagcaa cagcaggcag cagttcctca
     2941 gcagccactc gaccaattaa cacaatcaac tgcggaaact tggctctcca ttgcttcttt
     3001 ggcagaaacc cttggtgatg gcgacagggc cgcaatggca tatgacgcca ctttacagtt
     3061 caatccctca tctgcaaagg ctttaacatc cttggctcac ttgtaccgtt ccagagacat
     3121 gttccaaaga gctgcagaat tatatgaaag agcacttttg gtaaatcccg aactatcaga
     3181 tgtgtgggct actttaggtc attgttatct gatgctggat gatctgcaaa gagcttacaa
     3241 tgcctatcaa caggctctct accacctcag taatcccaac gtaccgaaat tatggcatgg
     3301 aatcggcatt ctttatgaca gatatggttc gctcgactat gccgaagaag cttttgccaa
     3361 agttttggaa ttggaccctc attttgaaaa ggcaaacgaa atttacttca gactaggtat
     3421 tatttataaa catcagggta aatggtctca agctttggaa tgcttcagat acattctccc
     3481 tcaacctcct gctcccttgc aggagtggga catatggttt cagttgggta gtgttttgga
     3541 gagtatggga gagtggcaag gtgcgaagga agcctacgag catgtcttgg ctcaaaatca
     3601 acatcatgcc aaagtattac aacaattagg ttgtctttac ggtatgagta acgtacaatt
     3661 ttatgaccct caaaaggcat tggattatct tctaaagtcg ttagaagcag atccctccga
     3721 tgccactaca tggtaccatc tcggtagagt gcatatgatt agaacagatt atactgccgc
     3781 atatgatgct ttccaacaag ctgttaatag agattcaaga aaccctatct tttggtgctc
     3841 aatcggtgtt ttatattacc aaatttctca atacagagac gccttagacg cgtacacaag
     3901 agccataaga ttaaatcctt atattagtga agtttggtac gatctaggta ctctttacga
     3961 aacttgtaac aaccaattat ctgacgccct tgatgcgtat aagcaagctg caagactgga
     4021 cgtaaataat gttcacataa gagaaagatt agaagcttta acaaagcagt tagaaaaccc
     4081 aggcaatata aacaaatcga acggtgcgcc aacgaatgcc tctcctgccc cacctcctgt
     4141 gattttacaa cctaccttac aacctaatga tcaaggaaat cctttgaaca ctagaatttc
     4201 agcccaatct gccaatgcta ctgcttcaat ggtacaacaa cagcatcctg ctcaacaaac
     4261 gcctattaac tcttctgcaa caatgtacag taatggagct tcccctcaat tacaagctca
     4321 agctcaagct caagctcaag cacaagctca agcacaagca caagctcaag cacaagcaca
     4381 agcacaagcg caagcacaag cacaagcaca ggcgcaagca caggcacaag cacaagcaca
     4441 agcacatgca caagcgcaag cacaagcaca agcacaggca caagcacaag cacaggcgca
     4501 ggcacaacaa caacaacaac aacaacaaca acaacaacaa caacaacaac aacaacaaca
     4561 acaacaacaa caacaacaac aacaacagca gcagcaatta cagcccctac caagacaaca
     4621 gctgcagcaa aagggagttt ctgtgcaaat gttaaatcct caacaagggc aaccatatat
     4681 cacacagcca acagtcatac aagctcacca actgcaacca ttttctacac aagctatgga
     4741 acatccgcaa agctctcaac tgccacctca acagcaacaa ctacaatctg ttcaacatcc
     4801 acaacaactt caaggccagc ctcaagccca agctccccaa cctttaatcc agcataacgt
     4861 ggaacagaac gttttacctc aaaagagata catggaaggt gcaatccaca ctttagtaga
     4921 tgccgccgta tccagtagca cccacacaga gaataacaca aagtctcctc gtcaaccaac
     4981 ccatgccatt ccaacgcaag ctcccgcaac aggaataacg aacgctgaac cacaggtaaa
     5041 gaagcaaaag ttgaactctc caaattcaaa catcaacaaa ttagtaaata ctgctacttc
     5101 cattgaagaa aatgcaaaat ctgaggtgag caaccaatcg ccagcagtag tggagtctaa
     5161 taccaataat acttcacaag aagaaaaacc tgtaaaagca aactcaatac cttcagtaat
     5221 tggcgcacag gaacctccac aggaagctag tcctgctgaa gaagctacca aagcagcttc
     5281 tgtttctcct tctacaaaac cgcttaatac ggaaccagag tcatctagtg tccaaccaac
     5341 tgtatcatca gaaagttcaa caacaaaagc aaatgaccaa agcactgctg agaccataga
     5401 actttctact gctactgttc ctgcagaagc aagccctgta gaagacgaag taagacagca
     5461 ttctaaagag gaaaacggca caactgaagc atctgcacct tctactgaag aggcggagcc
     5521 agcagcttcc agagatgctg aaaaacaaca agatgaaacc gctgctacaa cgataactgt
     5581 aatcaaacct actttggaaa caatggaaac agtgaaagag gaggccaaaa tgcgtgagga
     5641 agagcaaaca tctcaagaaa aatccccaca ggagaacaca cttccaagag aaaatgtagt
     5701 aaggcaagtg gaagaagatg aaaactacga cgactaaaaa gtttgcattc aaaaaattaa
     5761 tctactaatt tataatcaac gagaaatgtt gtgtagctca caattcattc attatgttgt
     5821 ggatacatat ataaataaag aaaaggctaa aaaaaaaggt gggtaacgtg aattaggtta
     5881 tttttcatat atgcacatgt aaatatatgc gtatataaat atattattat ttgtatcatt
     5941 acatacttaa aatctattat tgcttaaaag gttttcccga tgagcatatg taataatatt
     6001 gggaattaag gtgcattttc gtatccttca ttgtgtatct acaatctctt caagaaattc
     6061 ccttatttgc tttaaaacgg tttcccttgt tgaatccgat accatttcta gaaaaaaaaa
     6121 ttccactgtt agtaacgttc tcttttcaat aaatttagga tcccccgggc tgcaggaatt
     6181 cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtacca gcttttgttc
     6241 cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6301 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6361 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6481 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6541 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7081 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7381 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7741 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8401 tccgcgcaca tttccccgaa aagtgccacc tgggtccttt tcatcacgtg ctataaaaat
     8461 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     8521 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     8581 acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca
     8641 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     8701 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     8761 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     8821 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     8881 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     8941 gcgatccgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     9001 gaggcccttt cgtc
//
