LOCUS       sequence-193234-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Fluorescent CPI-17 neutral mutation 
            cytosolic.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  TITLE     Gordon Lab plasmids
REFERENCE   2  (bases 1 to 6536)
  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"
COMMENT     Sequence Label: DsRed-CPI-17-T38A-NES
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        22..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     650..669
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        650..668
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             688..1362
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(868..888)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1280..1303
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        complement(1876..1894)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1877..1894)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1914..1931)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1920..2144
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2190..2618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2277..2296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2487..2508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2627..2647)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2632..2961
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2812..2947
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2874..2893
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3028..3822
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVTCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3082..3101)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3692..3711
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3996..4117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4033..4052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4087..4106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4166..4182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4166..4182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4179..4201)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4190..4206
                     /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(join(4214..4220,4221..4238,4239..4244))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4259..4280
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4397..4414)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4568..5156)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4648..4667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5327..6118,6119..6187))
                     /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     5950..5969
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6188..6292)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6367..6386)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      661 actataggga gacccaagct tggtaccatg gcctcctccg agaacgtcat caccgagttc
      721 atgcgcttca aggtgcgcat ggagggcacc gtgaacggcc acgagttcga gatcgagggc
      781 gagggcgagg gccgccccta cgagggccac aacaccgtga agctgaaggt gaccaagggc
      841 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg
      901 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc
      961 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg cgaccgtgac ccaggactcc
     1021 tccctgcagg acggctgctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc
     1081 gacggccccg tgatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
     1141 ccccgcgacg gcgtgctgaa gggcgagacc cacaaggccc tgaagctgaa ggacggcggc
     1201 cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc
     1261 tactactacg tggacgccaa gctggacatc acctcccaca acgaggacta caccatcgtg
     1321 gagcagtacg agcgcaccga gggccgccac cacctgttcc tggaattcat ggcagctcag
     1381 cggctgggca agcgcgtgct gagcaagctg cagtctccat cgcgggcccg cgggccaggg
     1441 ggcagtcccg gggggctgca gaagcggcac gcgcgcgtcg ccgtcaagta tgaccggcgg
     1501 gagctgcagc ggcggctgga cgtggagaag tggatcgacg ggcgcctgga ggagctgtac
     1561 cgcggcatgg aggcagacat gcccgatgag atcaacattg atgaattgtt ggagttagag
     1621 agtgaagagg agagaagccg gaaaatccag ggactcctga agtcatgtgg gaaacctgtc
     1681 gaggacttca tccaggagct gctggcaaag cttcaaggcc tccacaggca gcccggcctc
     1741 cgccagccaa gcccctccca cgacggcagc ctcagccccc tccaggaccg ggcccggact
     1801 gctcaccccc tggccctgaa gctggccggc ctggacatcg gcagctgact cgagcatgca
     1861 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     1921 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1981 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2041 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2101 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2161 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2221 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2281 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2341 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2401 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2461 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2521 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2581 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2641 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2701 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2761 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2821 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2881 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     2941 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3001 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3061 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3121 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3181 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3241 acgacgggcg ttacttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3301 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3361 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3421 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3481 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3541 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3601 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3661 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3721 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     3781 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     3841 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     3901 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3961 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4021 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4081 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4141 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4201 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4261 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4321 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4381 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4441 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4501 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4561 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4621 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4681 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4741 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4801 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4861 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4921 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4981 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5041 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5101 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5161 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5221 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5281 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5341 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5401 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5461 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5521 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5581 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5641 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5701 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5761 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5821 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5881 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5941 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6001 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6061 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6121 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6181 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6241 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6301 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     6361 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     6421 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6481 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatg
//
