LOCUS       Exported                5837 bp ds-DNA     circular SYN 27-OCT-2021
DEFINITION  Mammalian expression of zebrafish Cavin4b(PRDmt)-mCherry. Parton lab
            clone JMD.
ACCESSION   .
VERSION     .
KEYWORDS    Cavin4b(PRDmt)-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5837)
  TITLE     Harriet's Cavin4 paper
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5837)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5837
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        77..1054
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1089..1107
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1089..1106
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1162..1186
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1187..1196
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(2148..2172)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2174..2881
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2316..2334)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2584..2603
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2778..2801
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(2883..2903)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        complement(2921..2938)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2943..3077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2967..2986)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3021..3040
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3190..3210)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3190..3208)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3229..3245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3229..3245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3242..3264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3253..3269
                     /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(3277..3307)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3322..3343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3460..3477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3631..4219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3711..3730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4390..5250)
                     /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     5013..5032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5251..5355)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5381..5836)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5518..5539)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5730..5749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg
      121 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
      181 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
      241 gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
      301 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
      361 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
      421 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
      481 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
      541 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
      601 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
      661 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
      721 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
      781 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
      841 ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg
      901 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
      961 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
     1021 ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag
     1081 caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg caggatccca
     1141 tcgattcgaa ttcaaggatc aacaagtttg tacaaaaaag caggctgcca ccatggctga
     1201 taaactggga ctggcaggtg tggcggacga gcccagcgcc atgaacatct tctctctgct
     1261 ggagagggtg tccgggatca tagacaatgt tcaggcctgc cagcagcgga tggaggagag
     1321 gcagctggag ctggagaaca ccgtcaagac catccaagca gatgtagtca agctgacgaa
     1381 ggagcacacg gcgaccagtg gcaccgtgga gaaacttctg gagaaaacac gcaaggtcag
     1441 ctgccacatc aaggacgtcc gcgtgcgggt ggagaagcag aacgttcgag tcaagaaggt
     1501 ggaggaaacc caagtcgagc tgctgaataa gaacaagttc cgtgtcgtca tctaccaggg
     1561 agataatgag gtcccagctg tggctggtcc taaagcctct ccgaagggag caatgggtgg
     1621 agacgctgct gcagacccag atgcaccaga ggctccgctc cctgactcgg atgaggaata
     1681 catggttgtt gaagaagccg attcttccac tgctgccaaa ctaaagaaga ccggcctgaa
     1741 gcgcatcgaa agcctgaagc agaccttctc acgcgagaac atgactaaga ccaaagaaaa
     1801 cctgggcacc aaggtcaaca agctgggcga gcgcatagta acagccgaac ggcgtgagaa
     1861 aatccgccag tcaggagaac gactgaaaca gtctggcgag cgcttcaagg aaaccatcac
     1921 aaaaaccgta ccagcaaagc tgaatctgaa gaaggaaagg acggtggctg agggtcagga
     1981 gggggcagaa gggaccacag aaggaacagc agccgtcaca ccaccgaagg gtcgtaaacc
     2041 cagtcctgag gtggcctaca cagagccaac gcagaaacaa gaaggtccag atgagggcaa
     2101 gggtgaggaa tcagaagtac cgatgtatga tatgaagcag ctttctgacc cagctttctt
     2161 gtacaaagtg gctgtgagca agggcgagga ggacaacatg gccatcatca aggagttcat
     2221 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     2281 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     2341 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     2401 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     2461 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     2521 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     2581 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     2641 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     2701 ctacgacgcc gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     2761 ctacaacgtc aacatcaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     2821 acagtacgag cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta
     2881 accaacttta ttatacatag ttgcactcga gcctctagaa ctatagtgag tcgtattacg
     2941 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3001 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3061 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3121 aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag
     3181 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     3241 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa
     3301 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3361 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3421 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3481 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3541 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3601 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3661 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3721 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3781 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3841 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3901 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3961 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4021 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4081 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4141 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4201 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4261 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4321 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4381 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4441 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4501 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4561 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4621 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4681 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4741 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4801 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4861 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4921 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4981 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5041 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5101 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5161 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5221 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5281 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5341 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     5401 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     5461 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     5521 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     5581 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     5641 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     5701 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     5761 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     5821 ccgctacagg gcgcgtc
//
