LOCUS       sequence-206489-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with 
            sarcoplasmic/endoplasmic calcium ATPase 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  AUTHORS   Hall TE, Martel N, Ariotti N, Xiong Z, Lo HP, Ferguson C, Rae J, Lim
            YW, Parton RG
  TITLE     In vivo cell biological screening identifies an endocytic capture 
            mechanism for T-tubule formation.
  JOURNAL   Nat Commun. 2020 Jul 24;11(1):3711. doi: 10.1038/s41467-020-17486-w.
  PUBMED    32709891
REFERENCE   2  (bases 1 to 5623)
  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: "Nat 
            Commun."; date: "2020-07-24"; volume: "11(1):3711. doi"; pages: " 
            10.1038/s41467-020-17486-w"
COMMENT     Sequence Label: p3E-SERCA1
FEATURES             Location/Qualifiers
     source          1..5623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1595..1604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3732..3827)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3864..3883)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3865..3883)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3888..3904)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3888..3904)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4006..4025)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4017..4826
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4084..4103)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4973..5561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5462..5481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggctg agaacgcaca caccaaggag acaccagaat
      781 gcctggccta cttcggcgtg agcgagtcca caggcctcac accagaacag gtcaagaaga
      841 accaagccaa gtacggattc aacgagctgc cagctgaaga aggtaagagc atctgggagc
      901 tggtggtgga gcagtttgaa gatctgttgg tcaggattct gcttttggcc gcctgcatct
      961 cttttgtact ggcctggttt gaggaaggtg aggaaacagt cactgccttc gtcgagccct
     1021 tcgttattct cctcatcctt atcgccaatg cagtggttgg agtatggcag gaacgcaatg
     1081 cggagagtgc cattgaggct ctgaaggaat atgagccaga gatgggcaaa gtctaccgct
     1141 ctgacagaaa gagtgtgcag aggatcaagg ccagagagat tgtccctgga gacattgtgg
     1201 aagtttcagt tggtgacaaa gtccctgctg atatcagaat caccgccatc cgctccacaa
     1261 ccctgcgcgt cgaccagtcc attcttactg gtgagtctgt tagtgtgatt aaacacacag
     1321 agtctgttcc tgaccccagg gctgtcaacc aggacaagaa gaacatgctt ttctctggca
     1381 ctaacatcgc tgctggcaag gctattggag ttgtggttgc cactggtgtg tccactgaaa
     1441 tcggtaaaat tcgtgaccag atggctgcca ctgagcagga gaagacccct ctgcagcaga
     1501 agctggatga gtttggtgag caactctcca aagtcatctc tctcatctgt gtggctgtct
     1561 ggctcatcaa catcggtcac ttcaacgacc ccgtccatgg tggctcctgg atccgtggtg
     1621 ccgtctacta cttcaagatc gctgtggctc ttgccgtagc tgccatcccc gaaggtttgc
     1681 ctgccgtcat cacaacctgc ttggctctgg gaacccgccg catggccaag aagaacgcca
     1741 ttgtccgctc tcttccctct gtagagactc tgggctgcac atctgtcatt tgctcagaca
     1801 agactggcac ccttaccacc aaccagatgt gtgtcacaaa gatgtttgtc attgagaaag
     1861 tggagggtga atctgtgact cttgaccagt acgacatttc tggatccaag tacacaccag
     1921 agggagaagt tactaagaac ggacttcctg tcaagtgcgg ccaatttgac ggccttgttg
     1981 agctggcaac catctgcgct ctgtgcaatg actcctctct ggactacaat gagtctaagg
     2041 gaatctatga gaaagttggc gaggctacag agactgccct ttgctgcctg gttgaaaaga
     2101 tgaatgtgtt caacactgat gttcgtggtc tgtccaaggt ggagagagcc aacacatgtt
     2161 gcgctgtgat caagcagctt atgaagaaag actttactct ggagttctcc cgtgacagga
     2221 agtccatgtc tgtctactgc tcccctgcaa aggcctccaa ggctcctgtt ggaaacaaga
     2281 tgtttgtcaa gggtgctcct gagggcgtta ttgacagatg tgcctatgtg cgtgttggca
     2341 ccactcgtgt gcccctgact ggtccagtta aagataagat catggctgtg atcaaggaat
     2401 ggggaactgg ccgtgatact ctgcgttgcc tggccctggc aacccgtgac aaccctctga
     2461 gaccagagga aatgaacttg gaagactcca ccaagtttgc tgaatatgag accgatctca
     2521 ccttcgtggg ttgtgttggt atgctggatc ctccccgtaa agaggttgtt ggatccattg
     2581 agctgtgcag ggctgctggc attcgtgtga tcatgatcac aggtgataac aagggcactg
     2641 ctgtggctat ctgccgccgt attggcatct tcagtgatga tgaggatgta acaggccgtg
     2701 ctttcactgg ccgtgagttt gatgacctgc cactgcccca gcagagagaa gcggtgcgta
     2761 aggcctgctg ttacgctcgt gttgagccct cccacaagag caagatcgtt gagttccttc
     2821 agggctttga tgagattact gccatgactg gtgatggtgt gaacgatgct cctgccctga
     2881 agaaggcaga gattggcatt gccatgggct ctggcactgc tgttgccaag tcagcctctg
     2941 agatggtcct ggccgacgac aacttctcta gcattgtggc cgctgttgag gagggcagag
     3001 ccatttacaa caacatgaag cagttcatcc gttacctcat ctcctctaat gttggagagg
     3061 ttgtctgtat tttcctgact gcagctcttg gtcttcctga ggctctgatc ccagttcagc
     3121 tgctgtgggt gaacctggtc actgatggtc tccctgccac tgctctgggc ttcaaccccc
     3181 ctgatttgga catcatgggc aaggctcccc gctcccccaa agagcctctt atctccggct
     3241 ggctcttctt cagatacctg gccattggag gttatgttgg tgctgcaact gttgctgctg
     3301 ctggatggtg gttcctctac tgtgatgaag gccccatggt ctccttctac cagctgtccc
     3361 acttcatgca gtgcactgct gacaatgagg actttgccgg tattgaatgt gaggtgttcg
     3421 aggccgctcc accaatgaca atggctctgt ctgtgttggt caccattgag atgtgcaatg
     3481 ctctgaacag cttgtcagag aatcagtctc ttttgcgtat gcctccatgg agcaacttgt
     3541 ggttggcggg agccatgact ctctccatgt ctctgcactt catgatcatc tatgtggacc
     3601 ctctgcctat gatcttcaaa ctaacccatc tcaactttga ccagtggatc gttgtactga
     3661 agctttcctt cccagtcatc ctcattgacg aagctctgaa atttgttgcc cgcaactacc
     3721 tggaggccta gcaactttat tatacaaagt tggcattata aaaaagcatt gcttatcaat
     3781 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgga gctccatggt
     3841 agcgttaacg cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     3901 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     3961 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     4021 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     4081 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     4141 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     4201 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     4261 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     4321 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     4381 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     4441 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     4501 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     4561 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     4621 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     4681 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     4741 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     4801 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     4861 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     4921 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4981 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5041 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5101 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5161 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5221 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5281 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5341 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     5401 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5461 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5521 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5581 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
