LOCUS       sequence-346599-        5548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Leishmania cell free expression of zebrafish mCherry-Bin1a. Parton 
            lab clone GSS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5548)
  AUTHORS   Lo HP, Lim YW, Xiong Z, Martel N, Ferguson C, Ariotti N, Giacomotto 
            J, Rae J, Floetenmeyer M, Moradi SV, Gao Y, Tillu VA, Xia D, Wang H,
            Rahnama S, Nixon SJ, Bastiani M, Day RD, Smith KA, Palpant NJ, 
            Johnston WA, Alexandrov K, Collins BM, Hall TE, Parton RG
  TITLE     Cavin4 interacts with Bin1 to promote T-tubule formation and 
            stability in developing skeletal muscle.
  JOURNAL   J Cell Biol. 2021 Dec 6;220(12). pii: 212693. doi: 
            10.1083/jcb.201905065. Epub 2021 Oct 11.
  PUBMED    34633413
REFERENCE   2  (bases 1 to 5548)
  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: "J Cell 
            Biol."; date: "2021-12-6"; volume: "220(12). pii: 212693. doi"; 
            pages: " 10.1083/jcb.201905065. Epub 2021 Oct 11"
COMMENT     Sequence Label: SITS-mCherry-Bin1a
FEATURES             Location/Qualifiers
     source          1..5548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    117..137
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     168..187
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        168..186
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             325..348
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     protein_bind    348..372
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      373..382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             379..1086
                     /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(524..542)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     792..811
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     986..1009
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(1088..1112)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2560..2580)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(2588..2604)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2620..2754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2644..2663)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2698..2717
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2919..2939)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2919..2937)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2958..2974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2958..2974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2971..2993)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2982..2998
                     /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(3006..3012,3013..3030,3031..3036))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3051..3072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3189..3206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3360..3948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3440..3459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4119..4910,4911..4979))
                     /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     4742..4761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4980..5084)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5110..5548,1..17))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5247..5268)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5459..5478
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
       61 ggtgcgggcc tcttcgctat tacgccagtc gactagggat aacagggtaa tgaattcaac
      121 tttgtataga aaagttgaca tttccccgaa aagtgccacc tgacgtctaa tacgactcac
      181 tatagggaca tcttaagttt attttatttt attttatttt attttatttt attttatttt
      241 attttatttt atttaaccat gacagtaatg tataaagtct gtaaagacat taaacacgta
      301 agtgaaacca tgggcacctc tggtcaccac catcaccacc atcaccacca agtttgtaca
      361 aaaaagcagg ctgccaccat ggtgagcaag ggcgaggagg acaacatggc catcatcaag
      421 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
      481 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
      541 aagggcggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
      601 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
      661 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
      721 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
      781 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
      841 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
      901 ggcggccact acgacgccga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
      961 cccggcgcct acaacgtcaa catcaagctg gacatcacct cccacaacga ggactacacc
     1021 atcgtggaac agtacgagcg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     1081 tacaagcacc cagctttctt gtacaaagtg gtggctgaat taccgatggg gaaaggggtg
     1141 tccgctggaa aaatagcaag caatgtacag aagaaaatta ctagagcaca ggagaaggtt
     1201 atgcaaaagc ttggaaaggc agatgagact aaagatctgg cgtttgaaga gggcgtgatc
     1261 aactttaata aacagttggt ggaaggtacc aaactgcaga aagaccttcg ggcctatttg
     1321 actgctgtga aagccatgca cgagtcttct agacgtctgt acgagtgcct ggatgacatg
     1381 tatgagacag agtggtacgg cagaaagaaa atggacgcag ttgtagagga ttcagacctg
     1441 ctttggacag acttccatca gaaactcgtg gatcatgcgc ttatatccat ggacacttac
     1501 ctgggccagt ttccagacat caaagctcgc attgcgaaac gtgacagaaa gctagttgac
     1561 tttgacagcg cacggcacaa ctttgctgct gtaaacaaag gcaagaagaa agaaggggtc
     1621 aagatcacca agccagcctc tttgctggaa agggctgccc caagctgggc tcaggggatc
     1681 ctgtccgccc ataatgtggc tcaaaccagc ctctccagga gtcaggcaga ggaggagctg
     1741 gagagggcac agaaagtctt tgaggaaatt aacatagatt tgcaggagga actgccatct
     1801 ctgtggaaca gtcgtgttgg cttttatgtg aatacgttcc agagtgtggc aggtctggag
     1861 gaaaagtttc acagagaaat gggaaaaatt aatcagaatc tttacgatac actggacaaa
     1921 ctggaaaatc aagatatgac cagtgaaaga acacaagatg ccaatcacac tttaagccca
     1981 ggaggaccac cagccattcc caaatcgccg tctaagctaa ggcctgcagt tcctcctcct
     2041 cccaaagtaa ctccgtcaaa ggacctgaag caggagagca tcatcaacct gtttgaagag
     2101 gccgccactc caaacatcag tgtcacttca ccaacagagt atgatgctcc agccactggt
     2161 aatcttctgg atatggatct ggactcactt caagctgcca ccacaccagc tgcacagacg
     2221 tgggattcat gggagactgc agaaacaaca gctgttacag aagaggcagc agcagctgaa
     2281 cctgtaagtg agactgtgca ggagagcgcc gagagcccag agagcgtaga gatgccagaa
     2341 tccttcctgt tcaaagtgaa agctatgcac gactacaatg ccaccgactc tgatgagctg
     2401 gagttaaagg cgggcgacgt ggtgctggta gtgaattatg aaaatccaga cgatcaggat
     2461 gacggctggt tgatgggagt caaagaggct gattggatcc agcagaaaga tcttgggaaa
     2521 aaaggcgtgt tccctgaaaa cttcacccag aggatgtgac aactttatta tacatagttg
     2581 ataattcact ggccgtcgtt ttacggtacc atcgatgatg atccagacat gataagatac
     2641 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2701 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2761 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2821 aagtaaaacc tctacaaatg tggtatggct gattatgatc tcgagattac cctgttatcc
     2881 ctagcgtgga tcctaggact agtacccagc ttttgttccc tttagtgagg gttaattgcg
     2941 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3001 ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3061 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3121 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3181 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3241 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3301 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3361 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3421 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3481 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3541 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3601 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3661 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3721 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3781 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3841 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3901 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3961 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4021 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4081 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4141 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4201 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4261 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4321 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4381 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4441 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4501 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4561 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4621 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4681 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4741 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4801 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4861 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4921 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4981 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5041 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5101 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5161 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5221 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5281 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5341 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5401 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5461 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5521 taaccaccac acccgccgcg cttaatgc
//
