LOCUS       sequence-193221-        6542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Fluorescent CPI-17 phospho mimetic nuclear.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6542)
  TITLE     Gordon Lab plasmids
REFERENCE   2  (bases 1 to 6542)
  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-NLS
FEATURES             Location/Qualifiers
     source          1..6542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     655..674
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        655..673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             693..1367
                     /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(873..893)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1285..1308
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1827..1847
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1887..1905)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1888..1905)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1925..1942)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1931..2155
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2201..2629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2288..2307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2498..2519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2638..2658)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2643..2972
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2823..2958
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2885..2904
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3039..3833
                     /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(3093..3112)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3703..3722
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4007..4128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4044..4063)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4098..4117
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4177..4193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4177..4193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4190..4212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4201..4217
                     /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(4225..4231,4232..4249,4250..4255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4270..4291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4408..4425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4579..5167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4659..4678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5338..6129,6130..6198))
                     /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     5961..5980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6199..6303)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6378..6397)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagaccc aagcttggta ccatggcctc ctccgagaac gtcatcaccg
      721 agttcatgcg cttcaaggtg cgcatggagg gcaccgtgaa cggccacgag ttcgagatcg
      781 agggcgaggg cgagggccgc ccctacgagg gccacaacac cgtgaagctg aaggtgacca
      841 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttccag tacggctcca
      901 aggtgtacgt gaagcacccc gccgacatcc ccgactacaa gaagctgtcc ttccccgagg
      961 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggcgacc gtgacccagg
     1021 actcctccct gcaggacggc tgcttcatct acaaggtgaa gttcatcggc gtgaacttcc
     1081 cctccgacgg ccccgtgatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc
     1141 tgtacccccg cgacggcgtg ctgaagggcg agacccacaa ggccctgaag ctgaaggacg
     1201 gcggccacta cctggtggag ttcaagtcca tctacatggc caagaagccc gtgcagctgc
     1261 ccggctacta ctacgtggac gccaagctgg acatcacctc ccacaacgag gactacacca
     1321 tcgtggagca gtacgagcgc accgagggcc gccaccacct gttcctggaa ttcatggcag
     1381 ctcagcggct gggcaagcgc gtgctgagca agctgcagtc tccatcgcgg gcccgcgggc
     1441 cagggggcag tcccgggggg ctgcagaagc ggcacgcgcg cgtcgaggtc aagtatgacc
     1501 ggcgggagct gcagcggcgg ctggacgtgg agaagtggat cgacgggcgc ctggaggagc
     1561 tgtaccgcgg catggaggca gacatgcccg atgagatcaa cattgatgaa ttgttggagt
     1621 tagagagtga agaggagaga agccggaaaa tccagggact cctgaagtca tgtgggaaac
     1681 ctgtcgagga cttcatccag gagctgctgg caaagcttca aggcctccac aggcagcccg
     1741 gcctccgcca gccaagcccc tcccacgacg gcagcctcag ccccctccag gaccgggccc
     1801 ggactgctca ccccggtgga ggtccaccta aaaagaagcg gaaagtgggt ggaggttgac
     1861 tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     1921 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1981 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2041 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2101 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2161 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2221 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2281 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2341 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2401 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2461 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2521 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2581 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2641 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2701 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2761 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2821 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2881 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     2941 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3001 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3061 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3121 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3181 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3241 cgtggctggc cacgacgggc gttacttgcg cagctgtgct cgacgttgtc actgaagcgg
     3301 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3361 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3421 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3481 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3541 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3601 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3661 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3721 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3781 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3841 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3901 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3961 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4021 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4081 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4141 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4201 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4261 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4321 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4381 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4441 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4501 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4981 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5281 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5401 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5461 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5521 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5581 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5641 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5701 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5761 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5821 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5881 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5941 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6001 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6061 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6301 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6361 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     6421 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     6481 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     6541 cg
//
