LOCUS       sequence-193212-        6500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Fluorescent CPI-17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6500)
  TITLE     Gordon Lab plasmids
REFERENCE   2  (bases 1 to 6500)
  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-wt
FEATURES             Location/Qualifiers
     source          1..6500
                     /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"
     promoter        complement(1845..1863)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1846..1863)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1883..1900)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1889..2113
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2159..2587
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2246..2265)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2456..2477
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2596..2616)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2601..2930
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2781..2916
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2843..2862
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2997..3791
                     /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(3051..3070)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3661..3680
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3965..4086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4002..4021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4056..4075
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4135..4151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4135..4151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4148..4170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4159..4175
                     /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(4183..4189,4190..4207,4208..4213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4228..4249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4366..4383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4537..5125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4617..4636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5296..6087,6088..6156))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHFTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5919..5938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6157..6261)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6336..6355)
                     /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 cgtcaccgtc 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 cccctgactc gagcatgcat ctagagggcc ctattctata gtgtcaccta
     1861 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1921 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1981 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2041 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2101 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2161 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2221 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2281 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2341 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2401 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2461 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2521 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2581 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2761 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2821 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2881 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2941 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3001 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3061 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3121 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3181 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tacttgcgca gctgtgctcg
     3241 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3301 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3361 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3421 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3481 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3541 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3601 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3661 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3721 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3781 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3841 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3901 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3961 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4081 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5821 cgtaaaatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6301 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6361 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6421 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     6481 aggcgttttg cgctgcttcg
//
