LOCUS       sequence-346578-        5194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Leishmania cell free expression of zebrafish mCherry-Bin1b. Parton 
            lab clone GSU.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5194)
  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 5194)
  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-Bin1b
FEATURES             Location/Qualifiers
     source          1..5194
                     /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(2206..2226)
                     /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(2234..2250)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2266..2400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2290..2309)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2344..2363
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2565..2585)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2565..2583)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2604..2620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2604..2620)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2617..2639)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2628..2644
                     /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(2652..2658,2659..2676,2677..2682))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2697..2718
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2835..2852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3006..3594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3086..3105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3765..4556,4557..4625))
                     /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     4388..4407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4626..4730)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(4756..5194,1..17))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4893..4914)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5105..5124
                     /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 gtggcagagg ttgggaaagg agtcaacgcc
     1141 gggaaactgg ccagtaatgt acagaagagg ttaacccgag cacaagagaa ggtgatgcag
     1201 aaactgggaa aggcggatga aaccaaagac acagcctttg aggagcaagt agccaacttt
     1261 aacaaacaga tgaatgaggg caccaaactg cagaaagact tgaaagcgta tttgctggca
     1321 gtcaaaacaa tgcacgagtg ttccaagcgt ctgcacgact gtctggcaga aatgtatgac
     1381 cctgagtggt ttggcaaaga ggaggtggac tccatcgcag aggacactga cctcctgtgg
     1441 caagatttcc acgagaagct tttggacaat gctgtaattt ccatggacgc atacttggct
     1501 cagtttccag atataaaggc tcgcattgca aagcgtgaaa ggaagctggt ggactttgac
     1561 agtgccagac atcattttgc ctctgtacaa aaaggcaaga aaaaggacga ggccaaaatt
     1621 gccaaggctg aggaacactt gggcagagct cagaaagtgt ttgaggagat caattatgat
     1681 ctccaggagg aacttcccac cctgtggaac agtcgtattg gcctctatgt taacacattc
     1741 cggagtgtgt ctggcctgga ggaaacattt cacagggaca tgggcaaact gaaccagaat
     1801 ctgagtgata ttatgaccaa actggaagag cagcgtgttg ccaaaaaagg tgtcacgaca
     1861 gcaagcactg ggaagagtga cgatgcagca aaccacaatc aggagacgtc agatgcacct
     1921 aaatctccat caaagaatga agccccaacg gtgacaaatg gctcctcaga tacagagctt
     1981 cctccatctg ttctctttaa ggtgaaaaca atgcacgact acagtgcaac agacagtgat
     2041 gaacttgacc tgaaggctgg tgacattgtg cttgtgcttc cctttgccaa cccagatgaa
     2101 gcggatgatg gctggttaat gggtgtaaaa gagtcccact ggcttcagaa caaaaacctc
     2161 ctagcaaaag gagtcttccc tgaaaacttc acccagaaac tgtgacaact ttattataca
     2221 tagttgataa ttcactggcc gtcgttttac ggtaccatcg atgatgatcc agacatgata
     2281 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2341 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2401 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2461 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatctcga gattaccctg
     2521 ttatccctag cgtggatcct aggactagta cccagctttt gttcccttta gtgagggtta
     2581 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2641 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     2701 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2761 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2821 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2881 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2941 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3001 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3061 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3121 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3181 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3241 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3301 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3361 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3421 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3481 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3541 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3601 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3661 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3721 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3781 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3841 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3901 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3961 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4021 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4081 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4141 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4201 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4261 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4321 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4381 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4441 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4501 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4561 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4621 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4681 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4741 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4801 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4861 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4921 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4981 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5041 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5101 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5161 tgcgcgtaac caccacaccc gccgcgctta atgc
//
