LOCUS       sequence-193220-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Fluorescent CPI-17 phospho mimetic 
            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-T38E-NES
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        35..414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        415..618
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     569..589
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     663..682
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        663..681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             701..1375
                     /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(881..901)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1293..1316
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        complement(1889..1907)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1890..1907)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1927..1944)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1933..2157
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2203..2631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2290..2309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2500..2521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2640..2660)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2645..2974
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2825..2960
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2887..2906
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3041..3835
                     /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(3095..3114)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3705..3724
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4009..4130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4046..4065)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4100..4119
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4179..4195)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4179..4195)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4192..4214)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4203..4219
                     /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(4227..4233,4234..4251,4252..4257))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4272..4293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4410..4427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4581..5169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4661..4680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5340..6131,6132..6200))
                     /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     5963..5982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6201..6305)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6380..6399)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
       61 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      121 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      181 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      241 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      301 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      361 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      421 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      481 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      541 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      601 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
      661 attaatacga ctcactatag ggagacccaa gcttggtacc atggcctcct ccgagaacgt
      721 catcaccgag ttcatgcgct tcaaggtgcg catggagggc accgtgaacg gccacgagtt
      781 cgagatcgag ggcgagggcg agggccgccc ctacgagggc cacaacaccg tgaagctgaa
      841 ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta
      901 cggctccaag gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt
      961 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggcgaccgt
     1021 gacccaggac tcctccctgc aggacggctg cttcatctac aaggtgaagt tcatcggcgt
     1081 gaacttcccc tccgacggcc ccgtgatgca gaagaagacc atgggctggg aggcctccac
     1141 cgagcgcctg tacccccgcg acggcgtgct gaagggcgag acccacaagg ccctgaagct
     1201 gaaggacggc ggccactacc tggtggagtt caagtccatc tacatggcca agaagcccgt
     1261 gcagctgccc ggctactact acgtggacgc caagctggac atcacctccc acaacgagga
     1321 ctacaccatc gtggagcagt acgagcgcac cgagggccgc caccacctgt tcctggaatt
     1381 catggcagct cagcggctgg gcaagcgcgt gctgagcaag ctgcagtctc catcgcgggc
     1441 ccgcgggcca gggggcagtc ccggggggct gcagaagcgg cacgcgcgcg tcgaggtcaa
     1501 gtatgaccgg cgggagctgc agcggcggct ggacgtggag aagtggatcg acgggcgcct
     1561 ggaggagctg taccgcggca tggaggcaga catgcccgat gagatcaaca ttgatgaatt
     1621 gttggagtta gagagtgaag aggagagaag ccggaaaatc cagggactcc tgaagtcatg
     1681 tgggaaacct gtcgaggact tcatccagga gctgctggca aagcttcaag gcctccacag
     1741 gcagcccggc ctccgccagc caagcccctc ccacgacggc agcctcagcc ccctccagga
     1801 ccgggcccgg actgctcacc ccctggccct gaagctggcc ggcctggaca tcggcagctg
     1861 actcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     1921 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1981 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2041 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2101 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2161 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2221 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2281 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2341 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2401 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2461 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2521 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2581 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2641 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2701 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2761 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2821 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2881 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     2941 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3001 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3061 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3121 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3181 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3241 atcgtggctg gccacgacgg gcgttacttg cgcagctgtg ctcgacgttg tcactgaagc
     3301 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3361 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3421 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3481 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3541 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3601 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3661 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3721 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3781 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3841 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3901 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3961 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4021 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4081 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4141 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4201 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4261 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4321 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4381 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4441 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4501 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4561 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4621 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4681 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4741 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4801 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4861 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4921 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4981 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5041 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5101 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5161 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5221 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5281 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5341 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5401 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5461 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5521 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5581 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5641 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5701 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5761 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5821 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5881 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5941 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6001 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6061 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6121 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6181 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6241 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6301 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     6361 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     6421 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     6481 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcg
//
