LOCUS       sequence-179932-        7466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses flag tagged mouse TBC1D13.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7466)
  AUTHORS   Davey JR, Humphrey SJ, Junutula JR, Mishra AK, Lambright DG, James 
            DE, Stockli J
  TITLE     TBC1D13 is a RAB35 specific GAP that plays an important role in 
            GLUT4 trafficking in adipocytes.
  JOURNAL   Traffic. 2012 Oct;13(10):1429-41. doi: 
            10.1111/j.1600-0854.2012.01397.x. Epub 2012 Jul 29.
  PUBMED    22762500
REFERENCE   2  (bases 1 to 7466)
  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: 
            "10.1111/j.1600-0854.2012.01397.x"; journalName: "Traffic"; date: 
            "2012-10"; volume: "13"; issue: "10"; pages: "1429-41"
COMMENT     Sequence Label: FLAG TBC1D13
FEATURES             Location/Qualifiers
     source          1..7466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(94..113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     304..325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        780..1159
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1160..1363
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1314..1334
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1393..1458
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2687..3309
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2886..2905)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3333..3353)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3351..3648
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3702..4496
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3756..3775)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4366..4385
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5148..5282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5185..5204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5239..5258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5287..5306)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5288..5306)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5320..5336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5320..5336)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5333..5355)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5344..5360
                     /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(5368..5374,5375..5392,5393..5398))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5413..5434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5551..5568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5722..6310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5802..5821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6481..7272,7273..7341))
                     /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     7104..7123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7342..7446)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
       61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      421 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg ccaagctgat ctatacattg aatcaatatt ggcaattagc
      661 catattagtc attggttata tagcataaat caatattggc tattggccat tgcatacgtt
      721 gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg
      781 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      841 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      901 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      961 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1021 agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     1081 gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt
     1141 agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg
     1201 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     1261 gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat
     1321 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     1381 gaattaacca tggactacaa agaccatgac ggtgattata aagatcatga catcgattac
     1441 aaggatgacg atgacaagct tgcggccgcg aattctatgt cgagtctgca caagagccgg
     1501 attgcagatt tccaggatgt cctgaaggag ccctccattg tgttggaaaa gcttcgagaa
     1561 ctcagtttta gtggcatccc ctgtgagggc ggactgcggt gcctctgctg gaagatcctc
     1621 ctgaactacc tccctctgga gagagcctcg tggacttcca tcctggccaa gcagaggggg
     1681 ctctattcgc agttcctgag agagatgatt atccagcctg gcattgccaa ggccaacatg
     1741 ggtgtattca gggaggatgt gacctttgag gaccatcctc tgaacccaaa ccctgacagc
     1801 aggtggaaca catattttaa ggacaacgag gtgctactcc agattgacaa agatgtcagg
     1861 agattgtgcc cagatatatc tttcttccaa agagccactg agtacccctg cctgctcatc
     1921 ctggacccgc aaaatgagtt tgagaccctt cgtaagcggg tggagcagac gacactgaag
     1981 tctcagacgg tggccaggaa tcgaagcggg gttacaaata tgagctcccc gcacaagaac
     2041 agtgcgccat cagccctgaa cgagtatgag gtgctaccca atggctgcga agcccactgg
     2101 gaggtggtgg agcgcatcct gttcatctat gccaagctca accctggcat cgcttacgtg
     2161 cagggcatga atgagatcgt gggccccctc tactacactt ttgccactga ccctaacagc
     2221 gagtggaaag agcacgctga agcagatacc tttttctgct tcaccaacct catggctgag
     2281 atccgggaca acttcatcaa gagcctggat gactcccagt gcggtatcac ctacaagatg
     2341 gaaaaggtgt actccacctt gaaggataag gatgtagagc tctacctgaa gctgcaagag
     2401 cagagtatca agcctcagtt ctttgccttc cgatggctga cattgctgct gtcccaggag
     2461 ttcttgctgc ctgatgtcat ccgtatctgg gactccctct ttgctgatgg caaccgattc
     2521 gatttcctcc tcctggtctg ctgtgctatg ctcatactga tccgggagca gttactggag
     2581 ggggacttca ctgtcaacat gcggcttctg caggattacc ccatcacaga cgtctgccag
     2641 atcctacaga aagccaagga actccaagac tcgaagtaag gatcccgggt ggcatccctg
     2701 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     2761 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     2821 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     2881 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     2941 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3001 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3061 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3121 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     3181 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     3241 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3301 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     3361 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3421 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3481 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3541 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3601 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gaattgatca
     3661 gcttgggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3721 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3781 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3841 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3901 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3961 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4021 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4081 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4141 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4201 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4261 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4321 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4381 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4441 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4501 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4561 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4621 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccaccccg ggctcgatcc
     4681 cctcgcgagt tggttcagct gctgcctgag gctggacgac ctcgcggagt tctaccggca
     4741 gtgcaaatcc gtcggcatcc aggaaaccag cagcggctat ccgcgcatcc atgcccccga
     4801 actgcaggag tggggaggca cgatggccgc tttggtcgac ccggacggga cgctcctgcg
     4861 cctgatacag aacgaattgc ttgcaggcat ctcatgagtg tgtcttcccg ttttctgcct
     4921 gaggtcactg cgtggatgga gcgctggcgc ctgctgcgcg acggcgagct gctcaccacc
     4981 cactcgccaa gctggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5041 cgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5101 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     5161 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5221 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     5281 tcaattccct atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg
     5341 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5401 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5461 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5521 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5581 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5641 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5701 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5761 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5821 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5881 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5941 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6001 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6061 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6121 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6181 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6241 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6301 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6361 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6421 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6481 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6541 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6601 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6661 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6721 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6781 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6841 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6901 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6961 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7021 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7081 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7141 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7201 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7261 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7321 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7381 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7441 tccgcgcaca tttccccgaa aagtgc
//
