LOCUS       sequence-311919-        7081 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Localization: Cilium, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7081)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 7081)
  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"
COMMENT     Sequence Label: mCherry-Smoothened1-N-18
FEATURES             Location/Qualifiers
     source          1..7081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     555..575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      633..642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3066..3075
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3072..3782
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3217..3235)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3485..3504
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3905..4026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3942..3961)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3996..4015
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4033..4488)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4170..4191)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4382..4401
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4515..4619
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4621..4978
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4621..4812
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4644..4664)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4829..4964
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5013..5807
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5013..5807
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5067..5086)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5677..5696
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5995..6014)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6039..6086
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6415..7003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6904..6923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
       61 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      421 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      541 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      601 agctggttta gtgaaccgtc agatccgcta gcgccaccat ggccgctggc cgccccgtgc
      661 gtgggcccga gctggcgccc cggaggctgc tgcagttgct tctgctggta ctgctggggg
      721 gcccgggccg gggggcggcc ttgagcggga acgtgaccgg gcctgggcct cacagcgcca
      781 gcgggagctc gaggagggac gtgccggtga ccagccctcc gccgccgctg ctgagccact
      841 gcggccgggc cgcccactgc gagcctttgc gctacaacgt gtgcctgggc tcggcgctgc
      901 cctacggagc caccaccacg ctgctggctg gggactcgga ctcgcaggag gaagcgcacg
      961 gcaagctcgt gctctggtcc ggcctccgga atgccccccg ctgctgggca gtgatccagc
     1021 ccctgctgtg tgctgtctac atgcccaagt gtgagaatga ccgagtggag ttgcccagcc
     1081 gtaccctctg ccaggccacc cgaggcccct gtgccattgt ggagcgggag cgagggtggc
     1141 ctgactttct gcgttgcaca ccggaccact tccctgaagg ctgcccaaac gaggtacaaa
     1201 acatcaagtt caacagctca ggccaatgtg aagcaccctt ggtgcgaaca gacaacccca
     1261 agagctggta tgaggacgtg gagggctgtg ggattcagtg tcagaacccg ctgttcaccg
     1321 aggccgagca ccaggacatg cacagctaca tcgcagcctt cggggcggtc actggtctct
     1381 gcacgctctt cacactggcc acctttgtgg ctgactggcg gaactccaat cgctaccctg
     1441 cggttattct cttctatgtc aatgcgtgtt tcttcgtggg cagcattggc tggctggccc
     1501 agttcatgga tggtgcccgc cgagagattg tttgccgagc agatggcacc atgagatttg
     1561 gggagcccac ctccagtgag accctgtcct gtgtcatcat ctttgtcatt gtgtactatg
     1621 ccttgatggc tggagtagtc tggttcgtgg tcctcaccta tgcctggcac acctccttca
     1681 aagccctggg caccacctac cagcctctct cgggcaagac atcctatttc cacctgctca
     1741 cgtggtcact cccctttgtc ctcacggtgg caatcctggc tgtggctcag gtagatggag
     1801 actccgtgag tggcatctgt tttgtaggct acaagaacta tcggtaccgt gctggctttg
     1861 tcctggcccc aattggcctg gtgcttattg tgggaggcta cttcctcatc agaggggtca
     1921 tgactctgtt ctccatcaag agcaaccacc ctgggcttct gagtgagaag gcagccagca
     1981 agatcaacga gaccatgctg cgcctgggca tttttggctt cctggccttt ggctttgtgc
     2041 tcatcacctt cagctgccac ttctatgact tcttcaacca ggctgagtgg gagcgtagct
     2101 tccgggacta tgtgctatgc caagccaacg tgaccatcgg gctgcctacc aagaagccca
     2161 ttcctgactg tgagatcaag aatcggccca gcctcctggt ggagaagatc aatctatttg
     2221 ccatgtttgg cactggcatt gccatgagca cctgggtctg gaccaaggcc accctgctca
     2281 tctggaggcg cacctggtgc aggttgactg ggcacagtga tgatgagccc aagagaatca
     2341 agaagagcaa gatgatcgcc aaggccttct ctaagcggcg tgagctgctg cagaacccgg
     2401 gccaggagct ctccttcagc atgcacactg tctcccatga tggacctgtt gcgggtttgg
     2461 cttttgacct caatgaaccc tcagctgacg tctcctctgc ctgggctcag catgtcacca
     2521 agatggtggc tcggagagga gccatattgc cccaggatgt gtccgttacc cctgtggcaa
     2581 ctccagtgcc accagaagaa caagccaaca tgtggctggt tgaggcagag atctccccag
     2641 agttagagaa gcgtttgggc cggaagaaaa agcggaggaa gaggaagaag gaggtgtgcc
     2701 ccttgaggcc agcccctgag cttcaccact ctgcccctgt tcctgccacc agtgcagttc
     2761 ctcggctgcc tcagctgcct cggcagaagt gcctggtagc tgcaaacgcc tggggaacag
     2821 gggagtcctg ccgacaggga gcctggactc tagtctccaa ccccttctgc ccagagccta
     2881 gtccccatca agatccattt ctccctggtg cctcagcccc ccgggtctgg gctcagggcc
     2941 gcctccaggg gctgggatcc attcattccc gcactaacct aatggaggct gagatcttgg
     3001 atgcagactc ggacttctct cgaattctgc agtcgacggt accgcgggcc cgggatccac
     3061 cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     3121 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     3181 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     3241 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     3301 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     3361 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     3421 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     3481 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     3541 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     3601 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     3661 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     3721 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     3781 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3841 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3901 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3961 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4021 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4081 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4141 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4201 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4261 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4321 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4381 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4441 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4501 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4561 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4621 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4681 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4741 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4801 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4861 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4921 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4981 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5041 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5101 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5161 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5221 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5281 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5341 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5401 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5461 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5521 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5581 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5641 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5701 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5761 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5821 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5881 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5941 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6001 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6061 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6121 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6181 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6241 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6301 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6361 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6421 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6481 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6541 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6601 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6661 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6721 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6781 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6841 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6901 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6961 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7021 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7081 c
//
