LOCUS       sequence-443324-        7576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ATP11B expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7576)
  AUTHORS   Gonzalez-Lozano MA, Schmid EW, Miguel Whelan E, Jiang Y, Paulo JA, 
            Walter JC, Harper JW
  TITLE     EndoMAP.v1 charts the structural landscape of human early endosome 
            complexes.
  JOURNAL   Nature. 2025 Jul;643(8070):252-261. doi: 10.1038/s41586-025-09059-y.
            Epub 2025 May 28.
  PUBMED    40437099
REFERENCE   2  (bases 1 to 7576)
  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"; doi: 
            "10.1038/s41586-025-09059-y"; journalName: "Nature"; date: 
            "2025-07"; volume: "643"; issue: "8070"; pages: "252-261"
COMMENT     Sequence Label: pGCS-3xFlag-ATP11B
FEATURES             Location/Qualifiers
     source          1..7576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1143..1277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1180..1199)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1234..1253
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1307..1331
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4893..4917)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(4927..4950)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4954..4977)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4981..5004)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(5042..5060)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5043..5060)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     rep_origin      5960..6415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6047..6066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6257..6278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6441..6545
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6546..6614,6615..7406)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGASKGGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6764..6783)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccgg gcccaatgca ttggcgccgc ggccgcgaat
     1081 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1141 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1201 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1261 cttatcatgt ctggatctac gtaatacgac tcactatagt tctagaacca ctttgtacaa
     1321 gaaagctggg cccatgaggc ctgaacaagt agatgagtcc attgtggaga gagtcaagat
     1381 gctcctgtcg ttggtataga aaggatccga tgcactccat gatctgctga tgtgggttgg
     1441 actacatctt cctataactc gttccagcat tcttccaaca gatgcacacg ctgcttctcc
     1501 ttccgggaaa cagcacatgg agtccaagca cttgatacct gcatttgttt cagtaagctg
     1561 tgccttttca gtacttgtag ggtggaggtg tcggtcaaag accttcttta tgatatcaag
     1621 aaatagacat gtaacaacca tgaggattat ggcaaaccaa gcagaaccac ttgacaggag
     1681 ctgaataaac acaaaataca tattctggga gcccaaaaat ggccagagaa tccctccata
     1741 aaacaaggaa aatacaaaat aaaatataat agatccccag gtaacgagat ggttgatcca
     1801 agtccaaaaa tgagtttcca gagccatctt tactgtgact gtaataacca tgactgtgaa
     1861 gaccaaagtg ccaaatgtcc agtttccaaa catctggcca tttccaagca gagatgtatc
     1921 tttccctatt agtaaatagg atccaaaaaa gaaaataaag gcatgactga agcccaggat
     1981 ggtccaataa agaaatgttt taatacttaa gaggcggttt ttactaatgt ctcgataaag
     2041 ggtgggctta ttttgtaaca catgagggtc tacatgctgt tccaaaagac tatatatcag
     2101 aataggtagg gaagtaaaac aaatattgta taaagtcagg tacacgctgt catacaatgt
     2161 ttgctgagaa aacaaacagt agaactgata taaaaactgg ggtgtgataa agcacacatt
     2221 cttataaaaa aaatactgta caagggtagc tattctaata taataaaaat gaccatgaac
     2281 aaaaagcaat ttggagagga acttaaatct ggctattgca tagtcactgt ttcttgcagc
     2341 ctgtcttcct tctttaccca tgattcctat gccaacatgg gcttcttgta tcatgcttac
     2401 gtcattagca ccatcaccaa cagccaatgt tataggtttc tcaggtgata tttttattag
     2461 tcttattact tttgctttct gcagtggagc catacgacag cataatacag ctgaacaatt
     2521 tctgcaaact tccataaata gtttttcatg ctccctgagt gcaagagata ggctggtccc
     2581 atccactacc agcccatgct gaatcacatg atcctctgta attcttctgg caagctgcct
     2641 caattgttca gcacactcgc tgtctgattt ctggtttata agttcaagga tgttcatggt
     2701 tctatgaaaa tggccacatg ataaactcac actaacagct gtttcatgtt tatccccagt
     2761 aagtacccat actttgatac cagccattct caatgcttca atagtttctc gaactttatc
     2821 ttgtagtctg tcttctactg ctgtggctcc aagtaatatc aggtctttct ctatgaactg
     2881 gaaaacagct gccaatttct cttcccgctg ctgcaaggca gtcctggctt caaatatgcg
     2941 tttatctatt tcctcatact cttttgatgt aaattttcta tatgctatac acagagttct
     3001 tagccctttc aaagcaaatt catctacatg aattctggtt ttttctattt ctccacctat
     3061 acatttaggg agaattgatg actcagctcc tttagcaaat aataacttct cacctgaagg
     3121 tgcctgaaca attacactca ttctcctacg atctgaatca aattccagaa tatgaagcag
     3181 tttgtaccgt tccagttttc caagagtttt aacctccata gtttcttcag aattgccaat
     3241 aaacacaata ccaatccttg cagcagcttc tactagagcc ttttcatctg gtgaagatgc
     3301 atagtactcc aactgcgatg gtgccaggtt ggattgccag ggaccatcac cagtgcagtc
     3361 agtttgaaca ttgctaatct gtacagtgtg acagagactg actgctttaa agaagagatc
     3421 atgttcttta attagttcag tttcattttc aggactggtt ctgaaagagg aactggttgt
     3481 aagatgggat aagttgttaa gatgggataa actactaaga taagataagt ttccttctga
     3541 agagtctggt gttggtcctt cgggtacaag tctaccatta atttcttggt atttcatgcc
     3601 attaattgaa cattcccgaa actgcatctc attttctgtc agtgtaccag ttttatctgt
     3661 aaacacgtac tctacctgtc caagctcttc attcagatcg gaagtattga cttgagcttt
     3721 ctgatctgat tcttcatgat acagatcaag atcccagcca ataaaaaatg atccaagaaa
     3781 tttctgcatt tcgactgtca catataatga aattggaatg atgaaattgt agagaaccaa
     3841 aaaagcaagg aagtctgaaa taaatctcag aatcttactg ctatttcttt gatgttctgt
     3901 tttttggtta taccaaggtt catcccattt ttcttcagct tgccatgtat acttcaagat
     3961 agtgctgatg acagcttcag atataagaat tactagataa attatcaaaa atgtattcat
     4021 tgacttttct actgcagatc gtttctgtga tttgctcttg taatttaatg ccatcttagt
     4081 ttccattcca gtgtataccg caacaccaaa aatttctttt gtgtttttta atctggctcc
     4141 acgaagcagg agactctccg gccccagagg tcttacaatt tcttccattt gttgggttat
     4201 gatcattcgt cccatgaatc tgtataagtc tgcttctggt tgctggcatt ctattacagc
     4261 tactagagtg tccaaattgg caactgtttg taataatgct gtttctggaa ctgccacatg
     4321 tgtcttcagg ttagtttctc cgtccaaact agcagttgta acgtgacagg aaccatccag
     4381 tcgatctgag gacagaagca ccaagtctgc aggaaaaatt tcatctttgg ctattcgaac
     4441 aatatcaccc acccgaatgt tttttgatct agtttttaca aggccaccac ttcgaacaac
     4501 ataaacagga gctccattta cttcattatc tgagttatgc cgtaaccaat cttcatatcc
     4561 ctgctttatg gcagttactg ttatcacaaa gaataatgga agtccactgg taactggact
     4621 ggtaggtgta tcaatcataa gctgaaccaa aaatataata agaaaataaa agtttgccac
     4681 tcttctgaac tgttcaaata aattttttgg aacaaaattc cacacagtgt acttagatga
     4741 aatgatcctg ttatctataa atttctgagg tgtgtaaagg ccattctgag gaaacctgtt
     4801 ggctacgtag atggttcttg tgtcactctg atgtggtggg tcaaaaccca gctgctgccg
     4861 gatccagcgc cacatggtgg gccttgacgg cccttctgct tttttgtaca aacttgttgg
     4921 ccggcctttg tcgtcatcat ctttataatc agccttatca tcgtcgtctt tgtagtctgc
     4981 tttgtcatcg tcatccttat aatccatggt ggatcctgca aaaagaacaa gtagcttgta
     5041 ttctatagtg tcacctaaat caagcttgct ccgacgtccc caggcagaat ggcggttccc
     5101 taaacgagca ttgcttatat agacctccca ttaggcacgc ctaccgccca tttacgtcaa
     5161 tggaacgccc atttgcgtca ttgcccctcc ccattgacgt caatggggat gtacttggca
     5221 gccatcgcgg gccatttacc gccattgacg tcaatgggag tactgccaat gtaccctggc
     5281 gtacttccaa tagtaatgta cttgccaagt tactattaat agatattgat gtactgccaa
     5341 gtgggccatt taccgtcatt gacgtcaata gggggcgtga gaacggatat gaatgggcaa
     5401 tgagccatcc cattgacgtc aatggtgggt ggtcctattg acgtcaatgg gcattgagcc
     5461 aggcgggcca tttaccgtaa ttgacgtcaa tgggggaggc gccatatacg tcaataggac
     5521 cgcccatatg acgtcaatag gaaagaccat atatagagac cattgacgtc aatgggggag
     5581 tggctatggg cggtattagg aagccccata tatggtatat gggaccgccc attgggaggg
     5641 gctatctacg tcaataggaa aacccatata tggaatacta tatggcatag ggccaataca
     5701 tagtattgaa cctggccaat agccatattg gcatggtgcc aagtccgcca tattgaattg
     5761 gctggggtcc agatccacca tattgaattg gcacgagtcc atatacgcca tattgaattg
     5821 gcacaggtcc agatccacca tattgaattg gcatgagtcc atatacgcca tattgaattg
     5881 gcggtcgact ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5941 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6001 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6061 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6121 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6181 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6241 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6301 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6361 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     6421 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6481 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6541 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6601 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6661 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6721 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6781 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6841 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6901 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6961 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7021 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7081 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7141 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7201 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc tccaagggtg
     7261 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7321 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7381 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7441 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7501 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7561 agatcaaagg atcttc
//
