LOCUS       Exported                4988 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    YIPlac204TKC-DsRed-Express2-HDEL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4988)
  TITLE     Glick lab plasmids
REFERENCE   2  (bases 1 to 4988)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4988
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      151..160
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             157..831
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(337..357)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     555..574
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     749..772
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     terminator      884..1131
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(900..918)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1150..1166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1150..1166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1163..1185)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1174..1190
                     /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(1198..1228)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1243..1264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1381..1398)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1552..2140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1632..1651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2311..3171)
                     /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     2934..2953
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3172..3276)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3344..3362
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3364..3465
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             3466..4140
                     /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"
     primer_bind     complement(4259..4281)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4381..4400
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgtttttca acagactaag cgctggcaag ctgctggtac cactctccgt ggtcctgtac
       61 gcccttttcg tggtaatatt acctttacag aattctttcc actcctccaa tgttttagtt
      121 agaggtgccg atgatgggga tccactagtc gccaccatgg atagcactga gaacgtcatc
      181 aagcccttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
      241 atcgagggcg agggcgaggg caagccctac gagggcaccc agaccgccaa gctgcaggtg
      301 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc
      361 tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc
      421 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
      481 caggactcct ccctgcagga cggcaccttc atctaccacg tgaagttcat cggcgtgaac
      541 ttcccctccg acggccccgt aatgcagaag aagactctgg gctgggagcc ctccaccgag
      601 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc acaaggcgct gaagctgaag
      661 ggcggcggcc actacctggt ggagttcaag tcaatctaca tggccaagaa gcccgtgaag
      721 ctgcccggct actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac
      781 accgtggtgg agcagtacga gcgcgccgag gcccgccacc acctgttcca gactcacggt
      841 atggacgaat tgtacaagca cgacgaattg taatctagag tcatcatgta attagttatg
      901 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
      961 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     1021 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     1081 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagcttggc
     1141 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1201 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1261 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1321 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1381 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1441 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1501 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1561 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1621 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1681 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1741 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1801 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1861 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1921 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1981 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2041 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2101 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2161 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2221 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2281 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2341 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2401 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2461 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2521 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2581 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2641 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2701 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2761 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2821 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2881 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2941 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3001 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3061 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3121 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3181 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3241 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3301 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3361 gccaattcgg tcgaaaaaag aaaaggagag ggccaagagg gagggcattg gtgactattg
     3421 agcacgtgag tatacgtgat taagcacaca aaggcagctt ggagtatgtc tgttattaat
     3481 ttcacaggta gttctggtcc attggtgaaa gtttgcggct tgcagagcac agaggccgca
     3541 gaatgtgcac tagattccga tgctgacttg ctgggtatta tatgtgtgcc caatagaaag
     3601 agaacaattg acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat
     3661 agttcaggca ctccgaaata cttggttggc gtgtttcgta atcaacctaa ggaggatgtt
     3721 ttggctctgg tcaatgatta cggcattgat atcgtccaac tgcatggaga tgagtcgtgg
     3781 caagaatacc aagagttcct cggtttgcca gttattaaaa gactcgtatt tccaaaagac
     3841 tgcaacatac tactcagtgc agcttcacag aaacctcatt cgtttattcc cttgtttgat
     3901 tcagaagcag gtgggacagg tgaacttttg gattggaact cgatttctga ctgggttgga
     3961 aggcaagaga gccccgagag cttacatttt atgttagctg gtggactgac gccagaaaat
     4021 gttggtgatg cgcttagatt aaatggcgtt attggtgttg atgtaagcgg aggtgtggag
     4081 acaaatggtg taaaagactc taacaaaata gcaaatttcg tcaaaaatgc taagaaatag
     4141 gttattactg agtagtattt atttaagtat tgtttgtgca cttgcctgca agccttttga
     4201 aaagcaagca taaaagatcg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     4261 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4321 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     4381 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     4441 gcgtaaggag aaaataccgc atcaggcgcc tacgttagtg tgagcgggat ttaaactgtg
     4501 aggaccttaa tacattcaga cacttctgcg gtatcaccct acttattccc ttcgagatta
     4561 tatctaggaa cccatcaggt tggtggaaga ttacccgttc taagactttt cagcttcctc
     4621 tattgatgtt acacctggac accccttttc tggcatccag tttttaatct tcagtggcat
     4681 gtgagattct ccgaaattaa ttaaagcaat cacacaattc tctcggatac cacctcggtt
     4741 gaaactgaca ggtggtttgt tacgcatgct aatgcaaagg agcctatata cctttggctc
     4801 ggctgctgta acagggaata taaagggcag cataatttag gagtttagtg aacttgcaac
     4861 atttactatt ttcccttctt acgtaaatat ttttcttttt aattctaaat caatcttttt
     4921 caattttttg tttgtattct tttcttgctt aaatctataa ctacaaaaaa cacatacata
     4981 aactaaaa
//
