LOCUS       sequence-210146-        6448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human rod opsin chimera with intracellular loop 3 from Scallop opsin
            with 1D4 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6448)
  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 6448)
  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: "BMC Biol.";
            date: "2018-01-16"; volume: "16(1):10. doi"; pages: " 
            10.1186/s12915-017-0475-2"
COMMENT     Sequence Label: pcDNA3 RL3Sc
FEATURES             Location/Qualifiers
     source          1..6448
                     /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(1805..1822)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1811..2035
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2081..2509
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2168..2187)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2378..2399
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2518..2538)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2523..2852
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2703..2838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2765..2784
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2919..3713
                     /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(2973..2992)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3583..3602
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3887..4008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3924..3943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3978..3997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4057..4073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4057..4073)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4070..4092)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4081..4097
                     /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(4105..4111,4112..4129,4130..4135))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4150..4171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4288..4305)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4459..5047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4539..4558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5218..6009,6010..6078))
                     /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     5841..5860
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6079..6183)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6258..6277)
                     /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 cgggcccacc ggctgcaacc
     1021 tggaaggctt ctttgctaca ctcggcggcg agattgctct gtggtcactg gtggtgctgg
     1081 ccatcgagag atacgtggtc gtgtgcaagc ccatgagcaa cttcagattc ggcgagaacc
     1141 acgccatcat gggcgtcgcc tttacatggg ttatggccct ggcttgtgcc gctcctccac
     1201 ttgctggctg gtccagatat atccctgagg gcctgcagtg cagctgcggc atcgattact
     1261 acaccctgaa gcctgaagtg aacaacgaga gcttcgtgat ctacatgttc gtggtgcact
     1321 tcacgatccc gatgatcatt atattctttt gctacggcca gctggtgttc accgtgaaag
     1381 tgtgcaagga cagccggaag aacggcatca gagcccagca gagatacacc cctcggttca
     1441 tccaggacat cgagcagcgc gtgaccttca tggtcattat tatggttatc gccttcctga
     1501 tctgctgggt gccctatgcc agcgtggcct tctacatctt cacccaccaa ggcagcaact
     1561 tcggccccat ctttatgaca atccccgcct tcttcgccaa gagcgccgcc atctacaacc
     1621 ccgtgatcta tatcatgatg aacaagcagt tccgcaactg catgctgacc accatctgct
     1681 gcggcaagaa tcctctggga gatgatgagg ccagcgccac cgtgtctaag accgagacat
     1741 ctcaggtggc ccctgcctaa gcggccgctc gagtctagag ggcccgttta aacccgctga
     1801 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1861 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1921 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1981 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2041 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2101 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2161 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2221 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2281 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2341 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2401 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2461 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2521 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2581 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2641 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2701 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2761 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     2821 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     2881 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2941 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3001 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3061 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3121 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3181 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3241 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3301 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3361 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3421 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3481 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3541 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3601 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3661 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3721 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3781 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3841 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3901 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3961 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4021 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4081 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4141 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4201 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4261 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4321 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4381 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4441 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4501 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4561 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4621 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4681 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4741 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4801 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4861 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4921 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4981 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5041 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5101 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5161 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5221 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5281 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5341 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5401 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5461 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5521 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5581 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5641 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5701 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5761 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5821 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5881 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5941 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6001 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6061 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6121 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6181 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6241 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     6301 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     6361 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     6421 cgcgatgtac gggccagata tacgcgtt
//
