pcDNA3 hML23Cm6

LOCUS       pcDNA3 hML23Cm6                6488 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Human melanopsin chimera with intracellular loops 2-3 and C-terminus
            from mGluR6.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3 hML23Cm6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6488)
  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 6488)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6488
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..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(1845..1862)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1851..2075
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2121..2549
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2208..2227)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2418..2439
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2558..2578)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2563..2892
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2743..2878
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2805..2824
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2959..3753
                     /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(3013..3032)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3623..3642
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3927..4048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3964..3983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4018..4037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4097..4113)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4097..4113)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4110..4132)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4121..4137
                     /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(4145..4175)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4190..4211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4328..4345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4499..5087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4579..4598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5258..6118)
                     /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     5881..5900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6119..6223)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6298..6317)
                     /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 gctagcgttt aaacttaagc ttgccaccat gaaccctcct tcggggccaa gagtcccgcc
      721 cagcccaacc caagagccca gctgcatggc caccccagca ccacccagct ggtgggacag
      781 ctcccagagc agcatctcca gcctgggccg gcttccatcc atcagtccca cagcacctgg
      841 gacttgggct gctgcctggg tccccctccc cacggttgat gttccagacc atgcccacta
      901 taccctgggc acagtgatct tgctggtggg actcacgggg atgctgggca acctgacggt
      961 catctatacc ttttgcagga gcagaagcct ccggacacct gccaacatgt tcattatcaa
     1021 cctcgcggtc agcgacttcc tcatgtcctt cacccaggcc cctgtcttct tcaccagtag
     1081 cctctataag cagtggctct ttggggagac aggctgcgag ttctatgcct tctgtggagc
     1141 tctctttggc atttcctcca tgatcaccct gacggccatc gccctggacc gtatctaccg
     1201 catctttgag cagggcaagc gctcggtcac accccctccc ttcatcagcc ccacctcaca
     1261 ggtcctgctg ggcgtttggc tctatgccct ggcctggagt ctgccaccct tcttcggctg
     1321 gagcgcctac gtgcccgagg ggttgctgac atcctgctcc tgggactaca tgagcttcac
     1381 gccggccgtg cgtgcctaca ccatgcttct ctgctgcttc gtgttcttcc tccctctgct
     1441 tatcatcatc tactgctaca tcttcatctt cagggcccgt ggcgtgcccg agaccttcaa
     1501 cgaggccaag atcatgctgc tggtcatcct cctcttcgtg ctctcctggg ctccctattc
     1561 cgctgtggcc ctggtggcct ttgctgggta cgcacacgtc ctgacaccct acatgagctc
     1621 ggtgccagcc gtcatcgcca aggcctctgc aatccacaac cccatcattt acgccatcac
     1681 ccaccccgag cagaatgtgc agaagcgaaa gcggagcctc aaggccacct ccacggtggc
     1741 agccccaccc aagggcgagg atgcagaggc ccacaagacg gagactagcc aggtggcccc
     1801 ggcctagctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     1861 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1921 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1981 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2041 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2101 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2161 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2221 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2281 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2341 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2401 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2461 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2521 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2581 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2641 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2701 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2761 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2821 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2881 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     2941 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3001 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3061 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3121 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3181 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3241 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3301 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3361 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3421 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3481 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3541 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3601 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3661 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3721 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3781 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3841 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3901 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     3961 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4021 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4081 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4141 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4201 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4261 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4321 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4381 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4441 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4501 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4561 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4621 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4681 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4741 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4801 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4861 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4921 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4981 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5041 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5101 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5161 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5221 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5281 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5341 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5401 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5461 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5521 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5581 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5641 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5701 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5761 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5821 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5881 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5941 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6001 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6061 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6121 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6181 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6241 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     6301 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     6361 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     6421 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     6481 tacgcgtt
//