LOCUS       Exported                8127 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses mKate2 under control of synthetic promoter containing 
            Zif268 binding sites..
ACCESSION   .
VERSION     .
KEYWORDS    pHES836
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8127)
  AUTHORS   Aranda-Diaz A, Mace K, Zuleta I, Harrigan P, El-Samad H
  TITLE     Robust Synthetic Circuits for Two-Dimensional Control of Gene 
            Expression in Yeast.
  JOURNAL   ACS Synth Biol. 2016 Dec 27. doi: 10.1021/acssynbio.6b00251.
  PUBMED    27930885
REFERENCE   2  (bases 1 to 8127)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8127
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        317..504
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     2589..2612
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2642..3340
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     primer_bind     complement(3342..3358)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3342..3358)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4677..4696)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4678..4696)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4703..4720)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4703..4719)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4712..4734)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4860..5315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4947..4966)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5157..5178
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5671..5690)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5790..5812
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5850..5868)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5953..6057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6058..6918
                     /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(6276..6295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7089..7677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7578..7597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7831..7848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7965..7986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8001..8031
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8039..8055
                     /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     8044..8066
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8063..8079
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8063..8079
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8100..8118
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctgggccggc catttaaatg tttaaacggc cacgctggcc gctgcacggt cctgttccct
       61 agcatgtacg tgagcgtatt tccttttaaa ccacgacgct ttgtcttcat tcaacgtttc
      121 ccattgtttt tttctactat tgctttgctg tgggaaaaac ttatcgaaag atgacgactt
      181 tttcttaatt ctcgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc attatcacat
      421 aatgaattat acattatata aagtaatgtg atttcttcga agaatatact aaaaaatgag
      481 caggcaagat aaacgaaggc aaagaggctc tatatgtata taaagctgcg gatgccttct
      541 gatttaatct agttctagtt tacttcaggt ttacttaaag ttgagataat tgaagcaaag
      601 agaagcatcg agcaaaggat ggcgtttgtt aagagggtta cgcaggagac gaatatacag
      661 ctggcgctgg atcttgacgg tgggtctgtt tctgtacggg agagcatact gggcaaggaa
      721 tatgctagtg gtgatgggca gaccatccat gtgcacactg gagttgggtt tttggaccac
      781 atgttgactg cgctggcgaa gcatggcggg tggtctctga tcctggagtg tataggggac
      841 ttgcacatcg atgaccacca cactgttgag gactgtggga ttgcgctggg ccaagcgttc
      901 aaggaggcgc ttggctccgt gcgtggtatc aagaggttcg ggcatgggtt tgcaccactg
      961 gacgaggcgc tgagccgcgc tgtggttgac ttctccaata ggcctttcgc cgtggtggag
     1021 ctgggcctga agcgagaacg cataggccag ctatccacag agatgatccc gcacttcttg
     1081 gagagtttcg ccactgaggc gcgtatcacc atgcatgtgg actgtctgcg gggcaccaac
     1141 gaccaccacc gctccgaatc agctttcaag gcgctcgcca tcgccatcag agaggcaaga
     1201 acacctacgg gtcgcgatga cgttccatcc actaagggtg tcctagcata gaacacagcc
     1261 cacagctacc accatcaaca atatttatat agataacgta cacatagaaa tcacacaaac
     1321 agagtattta ttcttaacta catgaactac catcagaccg tctgggccac tatataatgt
     1381 gctattcata aacgtgatca ctctacgtag caggcgaccc caggtgaaaa tttttcagcg
     1441 agctgccaga ttgtcaggtg aaaaactgaa aaaaacttct gggcgatgag cttgtggcgg
     1501 gaaattaagt atataaagca gttagtttct tctgcttctt gtggttctgg gtattcttgt
     1561 aatatttgga ataatagaaa accaagacta ctgcctttct tttcatatta tagaggaaga
     1621 aatacgcacg aacacgatat agaggtaaat tttttgcaaa aagataccaa actataaaag
     1681 aaatagaaga ctaaactaca aaaaataaag aaagtaatac atatatccaa tatgactgag
     1741 ttagcagagc aagtttccat gcctgaaggg caacaaagac gtggtggtta cgttccacct
     1801 cacatccgta acggtggtgg cagaagaggc gattctgctc ctcgtggtgg tttcaacaac
     1861 ggtggcggcc gtggtggttt cttcagaaac aacggtgacc gtggtggatt cggctttggg
     1921 taccgggccc ttatattgaa ttttcaaaaa ttcttacttt ttttttggat ggacgcaaag
     1981 aagtttaata atcatattac atggcattac caccatatac atatccatat acatatccat
     2041 atctaatctt acttatatgt tgtggaaatg taaagagccc cattatctta gcctaaaaaa
     2101 accttctctt tggaactttc agtaatacgc ttaactgctc attgctatat tgaagtgcgg
     2161 ccgcgtgggc gtgcgtgggc gggcgtgggc gtgcgtgggc gggcgtgggc gtgcgtgggc
     2221 gtctagaccg tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctaat
     2281 gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt atgaagagga
     2341 aaaattggca gtaacctggc cccacaaacc ttcaaattaa cgaatcaaat taacaaccat
     2401 aggatgataa tgcgattagt tttttagcct tatttctggg gtaattaatc agcgaagcga
     2461 tgatttttga tctattaaca gatatataaa tggaaaagct gcataaccac tttaactaat
     2521 actttcaaca ttttcagttt gtattacttc ttattcaaat gtcataaaag tatcaacaaa
     2581 aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaactatac tcgagaataa
     2641 aatggtgagc gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt
     2701 gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
     2761 gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
     2821 taccagcttc atgtacggca gcaaaacctt catcaaccac acccagggca tccccgactt
     2881 ctttaagcag tccttccccg agggcttcac atgggagaga gtcaccacat acgaagacgg
     2941 gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt
     3001 caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg
     3061 ctgggaggcc tccaccgaga ccctgtaccc cgctgacggc ggcctggaag gcagagccga
     3121 catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttgaaga ccacatacag
     3181 atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg acagaagact
     3241 ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc
     3301 cagatactgc gacctcccta gcaaactggg gcacagataa ggatccacta gttctagaca
     3361 cctgcaacat gcgtgagtaa ataagcggcc gccaccgcgg tggagctcta agcaaatagc
     3421 taaattatat acgaattaat attatgatta agtgtttacg tgagtgcgat atttttatta
     3481 ctatcttata cagttgtata tactctataa aatgagttgt ctattaatta acgcgatgaa
     3541 tgctttctgg gtttacctct ccaacaactc tagtttactt ctcaatacat tcaattgtat
     3601 ttgatttgtc aatacttcat cattaatcaa ttctatagtt ttgtttttct cgtttatttc
     3661 caaatttaat gcatcaattt tattattcaa tttgtcgttg attttggtta atgattttat
     3721 ggtttgatct ctggcattga ttgtttgtgt tagtttttca ttattgataa ttaaattatt
     3781 taagttagtt atcaactcgg tgttttcaag tttcaagttt tcaatttctt tagagtttat
     3841 tagatttgtc aaagtttctg aattgcttga ttggtcctgt agaagagtat ttgttgttgt
     3901 ggataattga ttcaattttt gagacaattg ctggaaggcg ttgaaatatc tagcatcaat
     3961 ctcatggttt ttttcccgag agtctcgtag attcaattgt tttaatatat cttgggacca
     4021 ctcttgattt gaactcatgg aaattaaact gggtgttgtg ttgtggtgta atgattgtac
     4081 cccctttgct tataattgtg tggtgacacc gattatttaa agctgcagca tacgatatat
     4141 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
     4201 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
     4261 ccttttttct ttttgctttt tctttttttt tctcttgaac tccagaaaaa aaatataaaa
     4321 gagatggagg aacgggaaaa agttagttgt ggtgataggt ggcaagtggt attccgtaag
     4381 aacaacaaga aaagcatttc atattatggc tgaactgagc gaacaagtgc aaaatttaag
     4441 catcaacgac aacaacgaga atggttatgt tcctcctcac ttaagaggaa aaccaagaag
     4501 tgccagaaat aacagtagca actacaataa caacaacggc ggctacaacg gtggccgtgg
     4561 cggtggcagc ttctttagca acaaccgtcg tggtggttac ggcaacggtg gtttcttcgg
     4621 tggaaacaac ggtggcagcg gccggccatt taaatgttta aacggccacg ctggccccct
     4681 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4741 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4801 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     4861 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4921 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4981 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5041 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5101 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5161 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5221 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5281 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     5341 catctgtgcg gtatttcaca ccgcataggg taataactga tataattaaa ttgaagctct
     5401 aatttgtgag tttagtatta ggatctctac ccttggcgaa aagtcctctg ccaacaatga
     5461 tgatatctga tccaccactt acaacttcgt cgacggttct gtactgctga cccaatatgc
     5521 gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta
     5581 ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg
     5641 aattgaaaag gtggtatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5701 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5761 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5821 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5881 tgtcatgata ataatggttt cttagacgtc ggcgcgccga ggacgtcagg tggcactttt
     5941 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6001 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6061 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6121 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6181 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6241 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6301 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6361 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6421 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6481 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6541 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6601 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6661 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6721 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6781 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6841 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6901 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6961 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7021 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7081 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7141 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7201 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7261 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7321 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7381 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7441 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7501 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7561 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7621 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7681 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7741 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7801 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7861 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7921 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7981 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     8041 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcgga
     8101 attaaccctc actaaaggga acaaaag
//
