LOCUS       sequence-314172-        7903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of TBC1D15.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7903)
  AUTHORS   Peralta ER, Martin BC, Edinger AL
  TITLE     Differential effects of TBC1D15 and mammalian Vps39 on Rab7 
            activation state, lysosomal morphology, and growth factor 
            dependence.
  JOURNAL   J Biol Chem. 2010 May 28. 285(22):16814-21.
  PUBMED    20363736
REFERENCE   2  (bases 1 to 7903)
  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 Biol 
            Chem. 2010 May 28. 285(22):16814-21."
COMMENT     Sequence Label: pEF6-myc-TBC1D15
FEATURES             Location/Qualifiers
     source          1..7903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1196..1215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1297..1306
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1306..1335
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3429..3470
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3480..3497
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3520..3537)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3526..3750
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3796..4224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3883..3902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4093..4114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4233..4253)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4238..4567
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4418..4553
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4480..4499
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4615..4662
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4681..5079
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5237..5358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5274..5293)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5328..5347
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5407..5423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5407..5423)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5420..5442)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5431..5447
                     /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(5455..5461,5462..5479,5480..5485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5500..5521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5638..5655)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5798..6386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5878..5897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6557..7348,7349..7417))
                     /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     7180..7199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7418..7522)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7597..7616)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt gcccttggaa ttcgccgccg ccatggaaca aaaactcatc
     1321 tcagaagagg atctggcggc ggcgggtgtt gtgagcggga agattatata tgaacaagaa
     1381 ggagtatata ttcactcatc ttgcggaaag gcaaatgacc aagacagctt gatttcagga
     1441 atattgcgtg ttttagaaaa ggatgctgaa gtaatagtgg actggagacc attggatgat
     1501 gcattagatt cttccagtat tctgtgtgct ggaaaggact ccagctcagt cgtagagtgg
     1561 actcaggctc caaaggaaag agctcatcgg gggtcggacc agcagagcag ctatgaagcg
     1621 gagtgggaca tggttactac agtgtccttt aagaagaaac cacacaccaa tggagatgct
     1681 ccaggccata ggaacgggaa aagcaaatgg tcgttcctgt tcagcctggc agacctgaag
     1741 tcagtcaagc agagcaagga gggcatgggc tggtcgtacc tggtcttctg cctgaaggat
     1801 gacgtcatgc tgcctgccct gcacttccat caaggagaca gcaaactgct gatcgagtct
     1861 cttgagaaat acgtggtatt gtgtgagtca ccacaagata gtagaacact tctagtcaat
     1921 tgtcagaata agagtctctc acagtctttt gaaaatcttc ttgatgaacc agcatatggc
     1981 ttaacacaaa aaattaaaaa ggatccttat acagcaacca tggtaggatt ttccaaagtt
     2041 acaaactaca tttttgacag tttgcgaggc agcgatccct ctacacatca acgaccacct
     2101 tcagaaatgg cagattttct tagtgacgct atcccaggac tcaagataaa ccagcaagaa
     2161 gagccaggat ttgaagtcat cactagaatt gatttgggag agcgacctgt tgtgcaaagg
     2221 cgggagccag tgtccctgga agaatggaac aagagccttg accctgaagg aagactcgta
     2281 gccgtggaga gcatgaagca aaagatattc agagggggtc ttagtcattc attaagaaag
     2341 caagcatgga aatttctcct gggttatttt ccttgggata gtaccaaaga ggaaagaact
     2401 cagttacaaa aacaaaaaac tgatgagtac ttcaggatga aactgcagtg gaagtctgtc
     2461 agtgaggctc aggagaagag aaactcacgc ctgagagact acagaagtct gatcgagaaa
     2521 gatgtcaaca gaacagatcg aacgaacaag ttttatgaag gccaagataa cccagggctg
     2581 attctgcttc acgacatttt gatgacctac tgtatgtatg actttgattt aggatatgta
     2641 caaggaatga gcgacttact ttccccactt ttgtatgtga tggagaatga agtggatgcc
     2701 ttttggtgct ttgcctctta catggaccag atgcatcaaa attttgaaga acaaatgcag
     2761 ggcatgaaga cgcagttaat ccagttgagt accttactca ggttactgga cagtggattt
     2821 tgtagctatt tagaatccca ggactctgga tatctatatt tttgcttcag atggctttta
     2881 atcagattta aaagggaatt tagttttcta gatatacttc gattatggga ggtaatgtgg
     2941 actgaactgc cctgtaagaa ttttcatctt cttctgtgtt gtgccatcct ggaatcagaa
     3001 aagcaacaga taatggcaaa gcactatggc ttcaatgaaa tccttaagca catcaatgag
     3061 ttgtccatga agattgatgt ggaagatata ctctgtaaag cagaagcaat ttccctacag
     3121 atggcacaat gcaaggaatt gccacaagca gtttgtgaga tcctgggact tcaggacaat
     3181 gaaattacaa cgccagattc cgacactgat gaaaatgttg gcagcccttg tcctgtatct
     3241 gcgtttccaa gtagcacctt gcccatcctt gctgccagtg aagccaaaga tgacagtcca
     3301 acacagacat tagcatcccc gaatgcctgc agattaacac ctgcatgatc atttctcgag
     3361 ccaagggcaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgc
     3421 ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc
     3481 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3541 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3601 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3661 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3721 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3781 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3841 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3901 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3961 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4021 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4081 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4141 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4201 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4261 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4321 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4381 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4441 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4501 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4561 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     4621 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     4681 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     4741 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     4801 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     4861 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     4921 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     4981 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     5041 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca
     5101 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5161 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5221 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5281 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5341 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5401 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5461 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5521 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5581 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5641 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5701 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5761 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5821 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5881 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5941 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6001 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6061 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6121 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6181 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6241 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6301 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6361 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6421 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6481 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6541 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6601 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6661 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6721 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6781 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6841 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6901 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6961 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7021 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7081 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7141 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7201 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7261 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7321 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7381 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7441 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7501 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     7561 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     7621 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     7681 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     7741 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     7801 gactaggctt ttgcaaaaag ctttgcaaag atggataaag ttttaaacag agaggaatct
     7861 ttgcagctaa tggaccttct aggtcttgaa aggagtggga att
//
