LOCUS       sequence-210137-        6436 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Human rod opsin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6436)
  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 6436)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 Rod Opsin
FEATURES             Location/Qualifiers
     source          1..6436
                     /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(1793..1810)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1799..2023
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2069..2497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2156..2175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2366..2387
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2506..2526)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2511..2840
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2691..2826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2753..2772
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2907..3701
                     /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(2961..2980)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3571..3590
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3875..3996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3912..3931)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3966..3985
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4045..4061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4045..4061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4058..4080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4069..4085
                     /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(4093..4099,4100..4117,4118..4123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4138..4159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4276..4293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4447..5035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4527..4546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5206..5997,5998..6066))
                     /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     5829..5848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6067..6171)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6246..6265)
                     /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 accgtcaagg
     1381 aggccgctgc ccagcagcag gagtcagcca ccacacagaa ggcagagaag gaggtcaccc
     1441 gcatggtcat catcatggtc atcgctttcc tcatatgctg ggtgccctac gccagcgtgg
     1501 cattctacat cttcacccac cagggctcca acttcggtcc catcttcatg accatcccag
     1561 cgttctttgc caagagcgct gccatctaca accctgtcat ctatatcatg atgaacaagc
     1621 agttccggaa ctgcatgctc accaccatct gctgcggcaa gaacccactg ggtgacgatg
     1681 aggcctctgc taccgtgtcc aagacggaga cgagccaggt ggccccggcc taaatcacta
     1741 gtgaattcgc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     1801 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1861 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     1921 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     1981 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2041 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2101 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2161 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2221 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2281 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2341 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2401 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2461 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2521 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2581 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2641 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2701 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2761 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2821 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     2881 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2941 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3001 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3061 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3121 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3181 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3241 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3301 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3361 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3421 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3481 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3541 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3601 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3661 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3721 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3781 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3841 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3901 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3961 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4021 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4081 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4141 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4201 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4261 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4321 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4381 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4441 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4501 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4561 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4621 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4681 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4741 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4801 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4861 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4921 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4981 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5041 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5101 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5161 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5221 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5281 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5341 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5401 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5461 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5521 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5581 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5641 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5701 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5761 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5821 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5881 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5941 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6001 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6061 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6121 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6181 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     6241 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     6301 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     6361 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     6421 gccagatata cgcgtt
//
