LOCUS       sequence-51779-e        8194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8194)
  AUTHORS   Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA, 
            McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin 
            MZ
  TITLE     Improving FRET dynamic range with bright green and red fluorescent 
            proteins.
  JOURNAL   Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171.
  PUBMED    22961245
REFERENCE   2  (bases 1 to 8194)
  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"; journalName: "Nat 
            Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171."
COMMENT     Sequence Label: pcDNA3-Camui-CR
FEATURES             Location/Qualifiers
     source          1..8194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      857..866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(908..929)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1516..1537
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3017..3026
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3023..3733
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     promoter        complement(3746..3764)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3747..3764)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3784..3801)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3790..4014
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4060..4488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4147..4166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4357..4378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4497..4517)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4502..4832
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4683..4818
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4745..4764
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4899..5693
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4953..4972)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5563..5582
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5867..5988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5904..5923)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5958..5977
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6037..6053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6037..6053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6050..6072)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6061..6077
                     /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(6085..6091,6092..6109,6110..6115))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6130..6151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6268..6285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6439..7027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6519..6538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7198..7989,7990..8058))
                     /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     7821..7840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8059..8163)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      841 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      901 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga
      961 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1021 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcgtgg cctgcttcag
     1081 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1141 cgtccaggag cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt
     1201 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1261 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat
     1321 cacggccgac aagcagaaga acggcatcaa ggctaacttc aagatccgcc acaacgttga
     1381 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1441 cgtgctgctg cccgacaacc actacctgag ccatcagtcc gccctgagca aagaccccaa
     1501 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggac tcagatctcg
     1561 agctatggct accatcacct gcacccgatt cacggaagag taccagctct tcgaggaact
     1621 gggaaaggga gccttctccg tggtgcgcag gtgtgtgaag gtgctggctg gccaggagta
     1681 tgctgccaag attatcaaca ccaagaagct ctcagccaga gatcaccaga agttggaacg
     1741 cgaggcccgc atctgccgct tgttgaagca ccccaatatc gtccgactcc atgacagcat
     1801 ctccgaggag gggcaccact accttatctt cgatctggtc actggtgggg agctgttcga
     1861 agacattgtg gcccgggagt attacagtga ggctgatgcc agccactgta tccagcagat
     1921 cctggaggct gtgctacact gtcaccagat gggggtggtg catcgcgacc tgaagcctga
     1981 gaatctgttg ctggcttcga agctcaaggg tgctgcggtg aagctggcag actttggcct
     2041 ggccatagag gttgagggag agcagcaggc atggtttggg ttcgcaggga cacctggata
     2101 cctctcccca gaagtgctgc ggaaggaccc atacgggaag cctgtggacc tgtgggcctg
     2161 tggcgtcatc ctgtatatct tgctggttgg gtatccccca ttctgggatg aggaccagca
     2221 ccgcctgtac cagcagatca aagctggtgc ctacgatttc ccatcaccag aatgggacac
     2281 cgtcaccccg gaagccaagg atctgatcaa taagatgctg accatcaacc cgtccaaacg
     2341 catcacggcc gctgaggctc tcaagcaccc ctggatctcg caccgctcca ctgtggcctc
     2401 ctgcatgcac agacaggaga ccgtggactg cctgaagaag ttcaatgcca ggaggaaact
     2461 gaagggagcc atcctcacca ctatgctggc caccaggaac ttctccggag ggaagagtgg
     2521 aggaaacaag aagaatgatg gcgtgaagga atcctctgag agcaccaaca ccaccatcga
     2581 ggatgaagac accaaagtgc gcaaacagga aattatcaaa gtgacagagc agctgatcga
     2641 agccataagc aatggagact ttgaatccta cacgaagatg tgcgaccctg gaatgacagc
     2701 ctttgaaccg gaggccctgg ggaacctggt cgagggcctg gactttcatc gattctattt
     2761 tgaaaacctg tggtcccgga acagcaagcc cgtgcacacc accatcctga accctcacat
     2821 ccacctgatg ggtgacgagt cagcctgcat cgcctacatc cgcatcactc agtacctgga
     2881 tgcgggtggc atcccccgca cggcccagtc agaggagacc cgtgtctggc accgcaggga
     2941 tggaaaatgg cagatcgtcc acttccacag atctggggcg ccctccgtcc tgccccatgg
     3001 actagcgcta ccggtcgcca ccatggtgtc taagggcgaa gagctgatca aggaaaatat
     3061 gcgtatgaag gtggtcatgg aaggttcggt caacggccac caattcaaat gcacaggtga
     3121 aggagaaggc aatccgtaca tgggaactca aaccatgagg atcaaagtca tcgagggagg
     3181 acccctgcca tttgcctttg acattcttgc cacgtcgttc atgtatggca gccgtacttt
     3241 tatcaagtac ccgaaaggca ttcctgattt ctttaaacag tcctttcctg agggttttac
     3301 ttgggaaaga gttacgagat acgaagatgg tggagtcgtc accgtcatgc aggacaccag
     3361 ccttgaggat ggctgtctcg tttaccacgt ccaagtcaga ggggtaaact ttccctccaa
     3421 tggtcccgtg atgcagaaga agaccaaggg ttgggagcct aatacagaga tgatgtatcc
     3481 agcagatggt ggtctgaggg gatacactca tatggcactg aaagttgatg gtggtggcca
     3541 tctgtcttgc tctttcgtaa caacttacag gtcaaaaaag accgtcggga acatcaagat
     3601 gcccggtatc catgccgttg atcaccgcct ggaaaggtta gaggaaagtg acaatgaaat
     3661 gttcgtagta caacgcgaac acgcagttgc caagttcgcc gggcttggtg gtgggatgga
     3721 cgagctgtac aagtaagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     3781 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3841 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3901 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3961 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4021 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4081 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4141 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4201 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4261 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4321 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4381 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     4441 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4501 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa
     4561 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4621 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4681 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4741 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4801 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4861 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4921 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4981 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5041 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5101 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5161 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5221 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5281 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5341 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5401 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5461 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5521 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5581 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5641 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5701 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5761 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5821 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5881 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5941 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6001 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6061 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6121 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6181 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6241 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6301 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6361 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6421 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6481 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6541 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6601 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6661 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6721 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6781 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6841 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6901 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6961 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7021 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7081 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7141 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7201 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7261 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7321 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7381 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7441 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7501 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7561 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7621 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7681 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7741 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7801 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7861 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7921 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7981 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8041 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8161 gcgcacattt ccccgaaaag tgccacctga cgtc
//
