LOCUS       sequence-51780-e        7050 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 7050)
  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 7050)
  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: pCAGGS-Raichu-RhoA-CR
FEATURES             Location/Qualifiers
     source          1..7050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     complement(1776..1797)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3285..3995
                     /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"
     primer_bind     complement(4074..4093)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4139..4194
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4193..4212)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4553..4569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4553..4569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4566..4588)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4577..4593
                     /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(4601..4607,4608..4625,4626..4631))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4646..4667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4726..4921
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4772..4907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4927..5061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4964..4983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5018..5037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5129..5146)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5300..5888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5380..5399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6059..6850,6851..6919))
                     /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     6682..6701
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6920..7024)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttgttgt gctgtctcat cattttggca agaattcggc atggtgagca
     1741 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1801 acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac ggcaagctga
     1861 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1921 ccttcggcta cggcgtggcc tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1981 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct ttcaaggacg
     2041 acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2101 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2161 acaacttcaa cagccacaac gtctatatca cggccgacaa gcagaagaac ggcatcaagg
     2221 ctaacttcaa gatccgccac aacgttgagg acggcagcgt gcagctcgcc gaccactacc
     2281 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcc
     2341 atcagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2401 tcgtaaccgc cgccctcgag tggtccctgc tagagcagct gggcctggcc ggggcagacc
     2461 tggcggcccc cggggtacag cagcagctgg agctggagcg ggagcggctg cggcgggaaa
     2521 tccgcaagga gctgaagctg aaggagggtg ctgagaacct gcggcgggcc accactgacc
     2581 tgggccgcag cctgggcccc gtagagctgc tgctgcgggg ctcctcgcgc cgcctcgacc
     2641 tgctgcacca gcagctgcag gagctgcacg cccacgtgac cggaggtgga accggtggtg
     2701 gaggtaccat ggctgccatc cggaagaaac tggtgattgt tggtgatgga gcctgtggaa
     2761 agacatgctt gctcatagtc ttcagcaagg accagttccc agaggtgtat gtgcccacag
     2821 tgtttgagaa ctatgtggca gatatcgagg tggatggaaa gcaggtagag ttggctttgt
     2881 gggacacagc tgggcaggaa gattatgatc gcctgaggcc cctctcctac ccagataccg
     2941 atgttatact gatgtgtttt tccatcgaca gccctgatag tttagaaaac atcccagaaa
     3001 agtggacccc agaagtcaag catttctgtc ccaacgtgcc catcatcctg gttgggaata
     3061 agaaggatct tcggaatgat gagcacacaa ggcgggagct agccaagatg aagcaggagc
     3121 cggtgaaacc tgaagaaggc agagatatgg caaacaggat tggcgctttt gggtacatgg
     3181 agtgttcagc aaagaccaaa gatggagtga gagaggtttt tgaaatggct acgagagctg
     3241 ctctgcaagc tagacgtggg aagaaaaaat ctggtggcgg ccgcatggtg tctaagggcg
     3301 aagagctgat caaggaaaat atgcgtatga aggtggtcat ggaaggttcg gtcaacggcc
     3361 accaattcaa atgcacaggt gaaggagaag gcaatccgta catgggaact caaaccatga
     3421 ggatcaaagt catcgaggga ggacccctgc catttgcctt tgacattctt gccacgtcgt
     3481 tcatgtatgg cagccgtact tttatcaagt acccgaaagg cattcctgat ttctttaaac
     3541 agtcctttcc tgagggtttt acttgggaaa gagttacgag atacgaagat ggtggagtcg
     3601 tcaccgtcat gcaggacacc agccttgagg atggctgtct cgtttaccac gtccaagtca
     3661 gaggggtaaa ctttccctcc aatggtcccg tgatgcagaa gaagaccaag ggttgggagc
     3721 ctaatacaga gatgatgtat ccagcagatg gtggtctgag gggatacact catatggcac
     3781 tgaaagttga tggtggtggc catctgtctt gctctttcgt aacaacttac aggtcaaaaa
     3841 agaccgtcgg gaacatcaag atgcccggta tccatgccgt tgatcaccgc ctggaaaggt
     3901 tagaggaaag tgacaatgaa atgttcgtag tacaacgcga acacgcagtt gccaagttcg
     3961 ccgggcttgg tggtggcatg gacgagctgt acaagtctag aaagatgagc aaagatggta
     4021 aaaagaagaa aaagaagtca aagacaaagt gtgtaattat gtaagtcgac ggatcttttt
     4081 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     4141 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     4201 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     4261 gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta
     4321 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     4381 gatttttttt tatattttgt ttgtgttatt tttttcttta acatccctaa aattttcctt
     4441 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     4501 tcttctctta tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     4561 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4621 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     4741 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4801 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4861 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4921 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4981 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5041 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5101 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5161 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5221 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5281 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5341 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5401 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5461 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     5521 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5581 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5641 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5701 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5761 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5821 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5881 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5941 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6001 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6061 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6121 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6181 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6241 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6301 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6361 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6421 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6481 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6541 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6601 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6661 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6721 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6781 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6841 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6901 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6961 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7021 cgcgcacatt tccccgaaaa gtgccacctg
//
