LOCUS       sequence-9897-ex        7669 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 7669)
  AUTHORS   Lewis TL, Mao T, Svoboda K, Arnold DB
  TITLE     Myosin-dependent targeting of transmembrane proteins to neuronal 
            dendrites.
  JOURNAL   Nat Neurosci. 2009 May . 12(5):568-76.
  PUBMED    19377470
REFERENCE   2  (bases 1 to 7669)
  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 
            Neurosci. 2009 May . 12(5):568-76."
COMMENT     Sequence Label: CHR2-MBD
FEATURES             Location/Qualifiers
     source          1..7669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             40..966
                     /codon_start=1
                     /product="N-terminal fragment of channelrhodopsin-2 from 
                     the alga Chlamydomonas reinhardtii (Nagel et al., 2003)"
                     /label=ChR2
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      994..1003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1000..1002,1003..1005,1006..1716)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1045..1066)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1306..1325)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1653..1674
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          1874..2007
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1990..2009
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2016..2604
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2069..2089)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2487..2498)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2631..2648)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2637..2748
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2975..2994)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3040..3095
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3094..3113)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3454..3470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3467..3489)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3478..3494
                     /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(3502..3508,3509..3526,3527..3532))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3547..3568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3627..3822
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3673..3808
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3828..3962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3865..3884)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3919..3938
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4030..4047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4201..4789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4281..4300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4960..5751,5752..5820))
                     /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     5583..5602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5821..5925)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5955..6334
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6337..6612
                     /label=chicken beta-actin promoter
     intron          6613..7621
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7545..7567
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 gaattcggta ccgcgggccc gggatccacc ggtgctagca tggattatgg aggcgccctg
       61 agtgccgttg ggcgcgagct gctatttgta acgaacccag tagtcgtcaa tggctctgta
      121 cttgtgcctg aggaccagtg ttactgcgcg ggctggattg agtcgcgtgg cacaaacggt
      181 gcccaaacgg cgtcgaacgt gctgcaatgg cttgctgctg gcttctccat cctactgctt
      241 atgttttacg cctaccaaac atggaagtca acctgcggct gggaggagat ctatgtgtgc
      301 gctatcgaga tggtcaaggt gattctcgag ttcttcttcg agtttaagaa cccgtccatg
      361 ctgtatctag ccacaggcca ccgcgtccag tggttgcgtt acgccgagtg gcttctcacc
      421 tgcccggtca ttctcattca cctgtcaaac ctgacgggct tgtccaacga ctacagcagg
      481 cgcaccatgg gtctgcttgt gtctgatatt ggcacaattg tgtggggcgc cacttccgcc
      541 atggccaccg gatacgtcaa ggtcatcttc ttctgcctgg gtctgtgtta tggtgctaac
      601 acgttctttc acgctgccaa ggcctacatc gagggttacc acaccgtgcc gaagggccgg
      661 tgtcgccagg tggtgactgg catggcttgg ctcttcttcg tatcatgggg tatgttcccc
      721 atcctgttca tcctcggccc cgagggcttc ggcgtcctga gcgtgtacgg ctccaccgtc
      781 ggccacacca tcattgacct gatgtcgaag aactgctggg gtctgctcgg ccactacctg
      841 cgcgtgctga tccacgagca tatcctcatc cacggcgaca ttcgcaagac caccaaattg
      901 aacattggtg gcactgagat tgaggtcgag acgctggtgg aggacgaggc cgaggctggc
      961 gcggtcaaca agggcaccgg caagccaccg gtcgccacca tggtgagcaa gggcgaggag
     1021 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1081 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1141 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1201 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1261 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1321 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1381 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1441 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1501 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1561 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     1621 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1681 gccgggatca ctctcggcat ggacgagctg tacaagggaa gcggtagcgg tacgcgtgga
     1741 tcaggctctc gagatcaacc actaaacagc aagaagaaga agcgactgct tagtttccga
     1801 gatgtagatt tcgaggagga cagcgattga atgcattgat gtacataaag cggccccgga
     1861 ctcgaggccg caggtaagta tcaaggttac aagacaggtt taaggagacc acatagaaac
     1921 tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg
     1981 acatccactt tgcctttctc tccacaggtg tcgacaatca acctctggat tacaaaattt
     2041 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2101 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2161 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2221 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2281 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     2341 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     2401 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     2461 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     2521 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     2581 tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag cctcgactgt
     2641 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2701 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2761 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2821 agacaatagc aggccagctt ttgttccctt tagggttaat tgcgcgcttg gcgctaatca
     2881 tggtcatagc tgtttccact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     2941 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     3001 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     3061 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     3121 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gccatatgct
     3181 ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     3241 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt ttatattttg
     3301 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     3361 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc
     3421 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3481 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3541 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3601 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     3661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3721 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3781 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     3841 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3901 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3961 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4381 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4621 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5941 agtgccacct ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     6001 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     6061 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     6121 aacgccaata gggactttcc attgacgtca atgggtggac tatttacggt aaactgccca
     6181 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     6241 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6301 gtacatctac gtattagtca tcgctattac catgggtcga ggtgagcccc acgttctgct
     6361 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
     6421 tattttgtgc agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg
     6481 gggcgagggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
     6541 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
     6601 gcgcggcggg cgggagtcgc tgcgttgcct tcgccccgtg ccccgctccg cgccgcctcg
     6661 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     6721 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggctcgtttc ttttctgtgg
     6781 ctgcgtgaaa gccttaaagg gctccgggag ggccctttgt gcggggggga gcggctcggg
     6841 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggcccgcgct gcccggcggc
     6901 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcgtgtgcgc gaggggagcg
     6961 cggccggggg cggtgccccg cggtgcgggg gggctgcgag gggaacaaag gctgcgtgcg
     7021 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc
     7081 ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgtg
     7141 cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg
     7201 gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggccccggag
     7261 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
     7321 gagggcgcag ggacttcctt tgtcccaaat ctggcggagc cgaaatctgg gaggcgccgc
     7381 cgcaccccct ctagcgggcg cgggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     7441 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccatctccag cctcggggct
     7501 gccgcagggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     7561 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     7621 gctcctgggc aacgtgctgg ttgttgtgct gtctcatcat tttggcaaa
//
