LOCUS       sequence-393316-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4733)
  AUTHORS   Nguyen AW, Daugherty PS
  TITLE     Evolutionary optimization of fluorescent proteins for intracellular 
            FRET.
  JOURNAL   Nat Biotechnol. 2005 Mar;23(3):355-60. doi: 10.1038/nbt1066
  PUBMED    15696158
REFERENCE   2  (bases 1 to 4733)
  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"; doi: "10"; journalName: 
            "Nat Biotechnol"; date: "2005-03"; volume: "23"; issue: "3"; pages: 
            "355-60"
COMMENT     Sequence Label: CYPet-N1
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5..799
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5..799
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(59..78)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     669..688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(987..1006)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1031..1078
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1407..1995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1896..1915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2170..2473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2474..2677
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2628..2648
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2700..2780
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2782..2791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2788..2790,2791..2793,2794..3507)
                     /codon_start=1
                     /product="cyan fluorescent protein optimized for FRET with 
                     YPet (Nguyen and Daugherty, 2005)"
                     /label=CyPet
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFGGIVPILVELEGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSVMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKARHNITDGSVQLADHYQQNTPIGDGPVILPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3094..3113)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3630..3751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3667..3686)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3721..3740
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3758..4213)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3895..3916)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4107..4126
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4240..4344
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4346..4703
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4346..4537
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4369..4389)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4554..4689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
       61 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
      121 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
      181 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
      241 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
      301 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
      361 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
      421 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
      481 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
      541 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
      601 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
      661 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
      721 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
      781 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
      841 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
      901 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
      961 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     1021 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     1081 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     1141 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     1201 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     1261 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     1321 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     1381 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1441 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1501 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1561 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1621 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1681 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1741 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1801 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1861 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1921 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     1981 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2041 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2101 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     2161 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2221 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2281 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2341 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     2401 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     2461 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     2521 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     2581 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     2641 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
     2701 ctagcgctac cggactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg
     2761 cgggcccggg atccaccggt cgccaccatg gtgagcaagg gagaggaact gttcggcggc
     2821 atcgtgccca tcctggtgga gctggagggc gacgtgaacg gccacaagtt cagcgtgagc
     2881 ggcgagggcg agggcgacgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     2941 ggcaagctgc ccgtgccctg gcccaccctg gtgaccaccc tgacctgggg cgtgcagtgc
     3001 ttcagccggt accccgacca catgaagcag cacgacttct tcaagagcgt gatgcccgag
     3061 ggctacgtgc aggagcggac catcttcttc aaggacgacg gcaactacaa gacccgggcc
     3121 gaggtgaagt tcgagggcga caccctggtg aaccggatcg agctgaaggg catcgacttc
     3181 aaggaggacg gcaacatcct gggccacaag ctggagtaca actacatcag ccacaacgtg
     3241 tacatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaaggc ccggcacaac
     3301 atcaccgacg gcagcgtgca gctggccgac cactaccagc agaacacccc catcggcgac
     3361 ggccccgtga tcctgcccga caaccactac ctgagcaccc agagcgccct gagcaaggac
     3421 cccaacgaga agcgggacca catggtgctg ctggagttcg tgaccgccgc cggcatcacc
     3481 cacggcatgg acgaactgta caaataaagc ggccgcgact ctagatcata atcagccata
     3541 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3601 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3661 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3721 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     3781 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3841 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3901 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3961 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4021 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4081 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4141 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4201 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4261 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4321 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4381 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4441 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4501 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4561 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4621 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4681 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtt
//
