LOCUS       sequence-157192-        5390 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 590, Emission:
            649.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5390)
  AUTHORS   Wang L, Jackson WC, Steinbach PA, Tsien RY
  TITLE     Evolution of new nonantibody proteins via iterative somatic 
            hypermutation.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Nov 30;101(48):16745-9. Epub 2004 Nov
            19.
  PUBMED    15556995
REFERENCE   2  (bases 1 to 5390)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2004-11-30"; volume: "101(48)"; pages: 
            "16745-9. Epub 2004 Nov 19"
COMMENT     Sequence Label: mPlum-pBAD
FEATURES             Location/Qualifiers
     source          1..5390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..145
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(71..90)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     231..253
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     391..410
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(590..1468)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1495..1779
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1702..1721
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1806..1828
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1848..1865
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1869..1901
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             1914..1934
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1941..2621
                     /codon_start=1
                     /product="far-red monomeric derivative of DsRed fluorescent
                     protein (Wang et al., 2004)"
                     /label=mPlum
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK
                     MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGA"
     primer_bind     2342..2361
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2536..2559
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2667..2684)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2837..2923
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3015..3042
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3061..3152
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3153..3221,3222..4013)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3371..3390)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4055..4510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4142..4161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4352..4373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4621..5209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5110..5129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5363..5380
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatctggt
       61 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
      121 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
      181 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
      241 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagaagg agatggcgcc
      301 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
      361 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
      421 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg
      481 tttgacagct tatcatcgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc
      541 aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact
      601 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
      661 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
      721 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
      781 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
      841 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
      901 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
      961 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     1021 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     1081 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     1141 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     1201 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     1261 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     1321 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     1381 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     1441 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa
     1501 ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc
     1561 taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca
     1621 tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt
     1681 tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggatacta
     1741 cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagaa
     1801 ataattttgt ttaactttaa gaaggagata tacatatgcg gggttctcat catcatcatc
     1861 atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg tacgagaacc
     1921 tgtacttcca gggctcgagc atggtgagca agggcgagga ggtcatcaag gagttcatgc
     1981 gcttcaagga gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     2041 gcgagggccg cccctacgag ggcacccaga ccgccaggct gaaggtgacc aagggtggcc
     2101 ccctgccctt cgcctgggac atcctgtccc ctcagatcat gtacggctcc aaggcctacg
     2161 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     2221 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     2281 tgcaggacgg cgagttcatc tacaaggtga aggtgcgcgg caccaacttc ccctccgacg
     2341 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     2401 aggacggcgc cctgaagggc gagatgaaga tgaggctgag gctgaaggac ggcggccact
     2461 acgacgccga ggtcaagacc acctacatgg ccaagaagcc cgtgcagctg cccggcgcct
     2521 acaagaccga catcaagctg gacatcacct cccacaacga ggactacacc atcgtggaac
     2581 agtacgagcg cgccgagggc cgccactcca ccggcgccta agaattcgaa gcttggctgt
     2641 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     2701 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     2761 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     2821 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactgag cctttcgttt
     2881 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     2941 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     3001 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     3061 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3121 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3181 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3241 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3301 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3361 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     3421 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3481 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3541 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3601 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3661 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     3721 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3781 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3841 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3901 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3961 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4021 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc
     4081 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4141 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4201 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4261 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4321 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4381 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4441 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4501 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4561 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4621 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4681 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4741 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4801 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4861 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4921 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4981 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5041 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5101 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5161 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5221 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5281 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5341 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
