LOCUS       sequence-118415-        6833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  optical depolarization of mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6833)
  AUTHORS   Offenhaeusser A
  TITLE     ChR2opt_mKATE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6833)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA4_ChR2opt_mKATE
FEATURES             Location/Qualifiers
     source          1..6833
                     /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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      984..993
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             990..1916
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     primer_bind     complement(2844..2861)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2850..3074
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3120..3548
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3207..3226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3417..3438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3557..3577)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3562..3891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3742..3877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3939..3986
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4005..4379
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4509..4630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4546..4565)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4600..4619
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4679..4695)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4679..4695)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4692..4714)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4703..4719
                     /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(4727..4733,4734..4751,4752..4757))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4772..4793
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4910..4927)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5081..5666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5161..5180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5837..6628,6629..6697))
                     /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     6460..6479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6698..6802)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttgccacca tggactacgg cggagccctg agcgccgtgg
     1021 gcagagaact gctgttcgtg accaaccccg tggtggtcaa cggcagcgtg ctggtgcccg
     1081 aggaccagtg ttactgcgcc ggctggatcg agagcagagg cacaaacggc gcccagaccg
     1141 cctctaacgt gctgcagtgg ctggccgctg gcttcagcat cctgctgctg atgttctacg
     1201 cctaccagac ctggaagtct acctgcggct gggaagagat ctacgtgtgc gccatcgaga
     1261 tggtcaaagt gatcctcgag ttcttcttcg aatttaaaaa ccccagcatg ctgtacctgg
     1321 ccaccggcca cagagtccag tggctgagat acgccgagtg gctgctgacc tgccccgtga
     1381 tcctgatcag actgagcaac ctgaccggcc tgagcaacga ctacagcaga cggaccatgg
     1441 gcctgctggt gtccgacatc ggcaccatcg tgtggggcgc cacaagcgcc atggccacag
     1501 gctacgtgaa agtgatcttc ttctgcctgg gcctgtgcta cggcgccaac acattcttcc
     1561 acgccgccaa ggcctacatc gagggctacc acaccgtgcc caagggcaga tgcagacagg
     1621 tggtcaccgg catggcctgg ctgttcttcg tgtcctgggg catgttcccc atcctgttca
     1681 tcctgggccc cgagggcttc ggcgtgctgt ctgtgtacgg cagcaccgtg ggccacacca
     1741 tcatcgacct gatgagcaag aactgctggg gactgctggg ccactacctg agagtgctga
     1801 tccacgagca catcctgatc cacggcgata tcagaaagac caccaagctg aacatcggcg
     1861 gcaccgagat cgaggtggaa accctggtcg aggacgaggc cgaggctggc gctgtgacag
     1921 ccgtggccac aatcgtgtcc aagggcgagg aactgatcaa agaaaacatg cacatgaagc
     1981 tgtacatgga aggcaccgtg aacaaccacc acttcaagtg caccagcgag ggcgagggca
     2041 agccctacga gggcacccag accatgagaa tcaaggtggt ggaaggcggc cctctgccct
     2101 tcgccttcga catcctggcc accagcttta tgtacggctc caagaccttc atcaaccaca
     2161 cccagggcat ccccgatttc ttcaagcaga gcttccctga gggcttcacc tgggagagag
     2221 tgaccaccta cgaggacggc ggcgtgctga ccgccaccca ggatacaagc ctgcaggacg
     2281 gctgcctgat ctacaacgtg aagatcagag gcgtgaactt ccccagcaac ggccccgtga
     2341 tgcagaagaa aaccctgggc tgggaggcca gcaccgagat gctgtaccct gccgacggcg
     2401 gcctggaagg cagagccgat atggccctga agctggtgga cggcggacac ctgatctgca
     2461 acctgaaaac cacctacaga agcaagaagc ccgccaagaa cctgaagatg cccggcgtgt
     2521 actacgtgga cagaaggctg gaaagaatca aagaggccga caaagagaca tacgtcgagc
     2581 agcacgaggt ggccgtggcc agatactgcg acctgcctag caagctgggc catagaggcg
     2641 gcggaggctc tggcctgaga tctagagccc aggctagcaa cagcgccgtg gatggcacag
     2701 ctggccctgg ctctaccggc agcagattct gctacgagaa cgaagtgtga ggatccacta
     2761 gtccagtgtg gtggaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg
     2821 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2881 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2941 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3001 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3061 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3121 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3181 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3241 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3301 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3361 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3421 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3481 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3541 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3601 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3661 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3721 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3781 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3841 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3901 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
     3961 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca
     4021 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
     4081 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
     4141 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
     4201 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
     4261 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
     4321 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
     4381 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4441 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4501 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4561 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4621 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4681 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4741 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4801 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4861 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4921 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4981 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5041 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5101 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5161 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5221 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5281 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5341 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5401 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5461 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5521 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5581 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     5641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5701 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5761 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5821 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5881 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5941 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6001 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6061 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6121 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6181 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6241 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6301 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6361 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6421 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6481 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6541 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6601 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6661 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6721 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6781 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
