LOCUS       sequence-222016-        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"
     protein_bind    complement(1..124)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      972..981
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3109..3204)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3241..3260)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3242..3260)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3265..3281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3265..3281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3383..3402)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3394..4203
                     /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(3461..3480)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4350..4938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4839..4858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5092..5109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5268..5295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5387..5473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5522..5544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5536..5553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5537..5553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 catagtgact gcatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
       61 taatttaata tattgatatt tatatcattt tacgtttctc gttcaacttt cttgtacaaa
      121 gtggctgaga acgcacacac caaggagaca ccagaatgcc tggcctactt cggcgtgagc
      181 gagtccacag gcctcacacc agaacaggtc aagaagaacc aagccaagta cggattcaac
      241 gagctgccag ctgaagaagg taagagcatc tgggagctgg tggtggagca gtttgaagat
      301 ctgttggtca ggattctgct tttggccgcc tgcatctctt ttgtactggc ctggtttgag
      361 gaaggtgagg aaacagtcac tgccttcgtc gagcccttcg ttattctcct catccttatc
      421 gccaatgcag tggttggagt atggcaggaa cgcaatgcgg agagtgccat tgaggctctg
      481 aaggaatatg agccagagat gggcaaagtc taccgctctg acagaaagag tgtgcagagg
      541 atcaaggcca gagagattgt ccctggagac attgtggaag tttcagttgg tgacaaagtc
      601 cctgctgata tcagaatcac cgccatccgc tccacaaccc tgcgcgtcga ccagtccatt
      661 cttactggtg agtctgttag tgtgattaaa cacacagagt ctgttcctga ccccagggct
      721 gtcaaccagg acaagaagaa catgcttttc tctggcacta acatcgctgc tggcaaggct
      781 attggagttg tggttgccac tggtgtgtcc actgaaatcg gtaaaattcg tgaccagatg
      841 gctgccactg agcaggagaa gacccctctg cagcagaagc tggatgagtt tggtgagcaa
      901 ctctccaaag tcatctctct catctgtgtg gctgtctggc tcatcaacat cggtcacttc
      961 aacgaccccg tccatggtgg ctcctggatc cgtggtgccg tctactactt caagatcgct
     1021 gtggctcttg ccgtagctgc catccccgaa ggtttgcctg ccgtcatcac aacctgcttg
     1081 gctctgggaa cccgccgcat ggccaagaag aacgccattg tccgctctct tccctctgta
     1141 gagactctgg gctgcacatc tgtcatttgc tcagacaaga ctggcaccct taccaccaac
     1201 cagatgtgtg tcacaaagat gtttgtcatt gagaaagtgg agggtgaatc tgtgactctt
     1261 gaccagtacg acatttctgg atccaagtac acaccagagg gagaagttac taagaacgga
     1321 cttcctgtca agtgcggcca atttgacggc cttgttgagc tggcaaccat ctgcgctctg
     1381 tgcaatgact cctctctgga ctacaatgag tctaagggaa tctatgagaa agttggcgag
     1441 gctacagaga ctgccctttg ctgcctggtt gaaaagatga atgtgttcaa cactgatgtt
     1501 cgtggtctgt ccaaggtgga gagagccaac acatgttgcg ctgtgatcaa gcagcttatg
     1561 aagaaagact ttactctgga gttctcccgt gacaggaagt ccatgtctgt ctactgctcc
     1621 cctgcaaagg cctccaaggc tcctgttgga aacaagatgt ttgtcaaggg tgctcctgag
     1681 ggcgttattg acagatgtgc ctatgtgcgt gttggcacca ctcgtgtgcc cctgactggt
     1741 ccagttaaag ataagatcat ggctgtgatc aaggaatggg gaactggccg tgatactctg
     1801 cgttgcctgg ccctggcaac ccgtgacaac cctctgagac cagaggaaat gaacttggaa
     1861 gactccacca agtttgctga atatgagacc gatctcacct tcgtgggttg tgttggtatg
     1921 ctggatcctc cccgtaaaga ggttgttgga tccattgagc tgtgcagggc tgctggcatt
     1981 cgtgtgatca tgatcacagg tgataacaag ggcactgctg tggctatctg ccgccgtatt
     2041 ggcatcttca gtgatgatga ggatgtaaca ggccgtgctt tcactggccg tgagtttgat
     2101 gacctgccac tgccccagca gagagaagcg gtgcgtaagg cctgctgtta cgctcgtgtt
     2161 gagccctccc acaagagcaa gatcgttgag ttccttcagg gctttgatga gattactgcc
     2221 atgactggtg atggtgtgaa cgatgctcct gccctgaaga aggcagagat tggcattgcc
     2281 atgggctctg gcactgctgt tgccaagtca gcctctgaga tggtcctggc cgacgacaac
     2341 ttctctagca ttgtggccgc tgttgaggag ggcagagcca tttacaacaa catgaagcag
     2401 ttcatccgtt acctcatctc ctctaatgtt ggagaggttg tctgtatttt cctgactgca
     2461 gctcttggtc ttcctgaggc tctgatccca gttcagctgc tgtgggtgaa cctggtcact
     2521 gatggtctcc ctgccactgc tctgggcttc aacccccctg atttggacat catgggcaag
     2581 gctccccgct cccccaaaga gcctcttatc tccggctggc tcttcttcag atacctggcc
     2641 attggaggtt atgttggtgc tgcaactgtt gctgctgctg gatggtggtt cctctactgt
     2701 gatgaaggcc ccatggtctc cttctaccag ctgtcccact tcatgcagtg cactgctgac
     2761 aatgaggact ttgccggtat tgaatgtgag gtgttcgagg ccgctccacc aatgacaatg
     2821 gctctgtctg tgttggtcac cattgagatg tgcaatgctc tgaacagctt gtcagagaat
     2881 cagtctcttt tgcgtatgcc tccatggagc aacttgtggt tggcgggagc catgactctc
     2941 tccatgtctc tgcacttcat gatcatctat gtggaccctc tgcctatgat cttcaaacta
     3001 acccatctca actttgacca gtggatcgtt gtactgaagc tttccttccc agtcatcctc
     3061 attgacgaag ctctgaaatt tgttgcccgc aactacctgg aggcctagca actttattat
     3121 acaaagttgg cattataaaa aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     3181 tcagtcaaaa taaaatcatt atttggagct ccatggtagc gttaacgcgg ccgcgatatc
     3241 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     3301 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     3361 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     3421 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     3481 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     3541 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     3601 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     3661 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     3721 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     3781 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     3841 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     3901 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     3961 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     4021 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     4081 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     4141 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     4201 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     4261 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     4321 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4381 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4441 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4501 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4561 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4621 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4681 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4741 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4801 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4861 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4921 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4981 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5041 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5101 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     5161 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     5221 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     5281 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     5341 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     5401 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     5461 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     5521 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggccctg cagctctaga
     5581 gctcgaattc tacaggtcac taataccatc taagtagttg gtt
//
