LOCUS       sequence-210142-        6427 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Human rod opsin chimera with intracellular loop 3 replaced by 
            intracellular loop 2 of mGluR6 with 1D4 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6427)
  AUTHORS   Ballister ER, Rodgers J, Martial F, Lucas RJ
  TITLE     A live cell assay of GPCR coupling allows identification of 
            optogenetic tools for controlling Go and Gi signaling.
  JOURNAL   BMC Biol. 2018 Jan 16;16(1):10. doi: 10.1186/s12915-017-0475-2.
  PUBMED    29338718
REFERENCE   2  (bases 1 to 6427)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC Biol.";
            date: "2018-01-16"; volume: "16(1):10. doi"; pages: " 
            10.1186/s12915-017-0475-2"
COMMENT     Sequence Label: pcDNA3 RL3m6L2
FEATURES             Location/Qualifiers
     source          1..6427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1784..1801)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1790..2014
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2060..2488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2147..2166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2357..2378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2497..2517)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2502..2831
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2682..2817
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2744..2763
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2898..3692
                     /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(2952..2971)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3562..3581
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3866..3987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3903..3922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3957..3976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4036..4052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4036..4052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4049..4071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4060..4076
                     /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(4084..4090,4091..4108,4109..4114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4129..4150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4267..4284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4438..5026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4518..4537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5197..5988,5989..6057))
                     /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     5820..5839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6058..6162)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6237..6256)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gccagcgttt aaacttaagc ttcaccatga atggcacaga aggccctaac ttctacgtgc
      721 ccttctccaa tgcgacgggt gtggtacgca gccccttcga gtacccacag tactacctgg
      781 ctgagccatg gcagttctcc atgctggccg cctacatgtt tctgctgatc gtcctaggct
      841 tccccataaa tttcctcacg ctctacgtca ccgtccagca caagaagctg cgcacgcctc
      901 tcaactacat cctgctcaac ctagccgtgg cagatctctt catggttcta ggtggcttca
      961 ccagcaccct ctacacctct ctgcatggat acttcgtctt cgggcccaca ggatgcaatt
     1021 tggagggctt ctttgccacc ctgggcggtg aaattgccct gtggtccttg gtggtcctgg
     1081 ccatcgagcg gtacgtggtg gtgtgtaagc ccatgagcaa cttccgcttc ggggagaacc
     1141 atgccatcat gggcgttgcc ttcacgtggg tcatggcgct ggcctgcgcc gcacccccac
     1201 tcgccggctg gtccaggtac atccccgagg gcctgcagtg ctcgtgtgga atcgactact
     1261 acacgctcaa gccggaggtc aacaacgagt cttttgtcat ctacatgttc gtggtccact
     1321 tcaccatccc catgattatc atctttttct gctatgggca gctcgtcttc accgtccgta
     1381 tctaccgcat cttcgaacag ggcaagcgct cggtcacacc ccctcccttc atcagcccca
     1441 cctcacagat ggtcatcatc atggtcatcg ctttcctcat atgctgggtg ccctacgcca
     1501 gcgtggcatt ctacatcttc acccaccagg gctccaactt cggtcccatc ttcatgacca
     1561 tcccagcgtt ctttgccaag agcgctgcca tctacaaccc tgtcatctat atcatgatga
     1621 acaagcagtt ccggaactgc atgctcacca ccatctgctg cggcaagaac ccactgggtg
     1681 acgatgaggc ctctgctacc gtgtccaaga cggagacgag ccaggtggcc ccggcctaag
     1741 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     1801 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1861 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1921 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1981 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2041 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2101 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2161 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2221 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2281 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2341 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2401 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2461 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2581 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2641 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2701 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2761 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2821 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     2881 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2941 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3001 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3061 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3121 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3181 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3241 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3301 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3361 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3421 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3481 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3541 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3601 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3661 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3721 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3781 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3841 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3901 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3961 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4021 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4081 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4141 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4201 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4261 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4321 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4381 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4441 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4501 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4561 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4621 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4681 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4741 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4801 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4861 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4921 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4981 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5041 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5101 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5161 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5221 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5281 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5341 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5401 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5461 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5521 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5581 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5641 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5701 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5761 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5821 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5881 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5941 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6001 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6061 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6121 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6181 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     6241 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     6301 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     6361 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     6421 acgcgtt
//
