LOCUS       sequence-313779-        7143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7143)
  AUTHORS   Venkatachalam K, Hofmann T, Montell C
  TITLE     Lysosomal localization of TRPML3 depends on TRPML2 and the 
            mucolipidosis-associated protein TRPML1.
  JOURNAL   J Biol Chem. 2006 Jun 23. 281(25):17517-27.
  PUBMED    16606612
REFERENCE   2  (bases 1 to 7143)
  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. 2006 Jun 23. 281(25):17517-27."
COMMENT     Sequence Label: TRPML1-MYC
FEATURES             Location/Qualifiers
     source          1..7143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2433..2462
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2500..2517)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2506..2730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2776..3204
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2863..2882)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3073..3094
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3213..3233)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3218..3547
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3398..3533
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3460..3479
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3614..4408
                     /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(3668..3687)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4278..4297
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4582..4703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4619..4638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4673..4692
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4752..4768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4752..4768)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4765..4787)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4776..4792
                     /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(4800..4806,4807..4824,4825..4830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4845..4866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4983..5000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5154..5742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5234..5253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5913..6704,6705..6773))
                     /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     6536..6555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6774..6878)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6953..6972)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttaccagcca ccatgacagc cccggcgggt ccgcgcggct
      721 cagagaccga gcggcttctg acccccaacc ccgggtatgg gacccaggcg gggccttcac
      781 cggcccctcc gacaccccca gaagaggaag accttcgccg tcgtctcaaa tactttttca
      841 tgagtccctg cgacaagttt cgagccaagg gccgcaagcc ctgcaagctg atgctgcaag
      901 tggtcaagat cctggtggtc acggtgcagc tcatcctgtt tgggctcagt aatcagctgg
      961 ctgtgacatt ccgggaagag aacaccatcg ccttccgaca cctcttcctg ctgggctact
     1021 cggacggagc ggatgacacc ttcgcagcct acacgcggga gcagctgtac caggccatct
     1081 tccatgctgt ggaccagtac ctggcgttgc ctgacgtgtc actgggccgg tatgcgtatg
     1141 tccgtggtgg gggtgaccct tggaccaatg gctcagggct tgctctctgc cagcggtact
     1201 accaccgagg ccacgtggac ccggccaacg acacatttga cattgatccg atggtggtta
     1261 ctgactgcat ccaggtggat ccccccgagc ggccccctcc gccccccagc gacgatctca
     1321 ccctcttgga aagcagctcc agttacaaga acctcacgct caaattccac aagctggtca
     1381 atgtcaccat ccacttccgg ctgaagacca ttaacctcca gagcctcatc aataatgaga
     1441 tcccggactg ctataccttc agcgtcctga tcacgtttga caacaaagca cacagtgggc
     1501 ggatccccat cagcctggag acccaggccc acatccagga gtgtaagcac cccagtgtct
     1561 tccagcacgg agacaacagc ttccggctcc tgtttgacgt ggtggtcatc ctcacctgct
     1621 ccctgtcctt cctcctctgc gcccgctcac tccttcgagg cttcctgctg cagaacgagt
     1681 ttgtggggtt catgtggcgg cagcggggac gggtcatcag cctgtgggag cggctggaat
     1741 ttgtcaatgg ctggtacatc ctgctcgtca ccagcgatgt gctcaccatc tcgggcacca
     1801 tcatgaagat cggcatcgag gccaagaact tggcgagcta cgacgtctgc agcatcctcc
     1861 tgggcacctc gacgctgctg gtgtgggtgg gcgtgatccg ctacctgacc ttcttccaca
     1921 actacaatat cctcatcgcc acactgcggg tggccctgcc cagcgtcatg cgcttctgct
     1981 gctgcgtggc tgtcatctac ctgggctact gcttctgtgg ctggatcgtg ctggggccct
     2041 atcatgtgaa gttccgctca ctctccatgg tgtctgagtg cctgttctcg ctcatcaatg
     2101 gggacgacat gtttgtgacg ttcgccgcca tgcaggcgca gcagggccgc agcagcctgg
     2161 tgtggctctt ctcccagctc tacctttact ccttcatcag cctcttcatc tacatggtgc
     2221 tcagcctctt catcgcgctc atcaccggcg cctacgacac catcaagcat cccggcggcg
     2281 caggcgcaga ggagagcgag ctgcaggcct acatcgcaca gtgccaggac agccccacct
     2341 ccggcaagtt ccgccgcggg agcggctcgg cctgcagcct tctctgctgc tgcggaaggg
     2401 acccctcgga ggagcattcg ctgctggtga atgaacaaaa attaataagt gaagaagatt
     2461 tactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     2521 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2581 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2641 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2701 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     2761 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2821 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2881 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2941 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3001 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3061 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3121 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3181 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3361 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3421 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3481 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3541 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3601 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3661 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3721 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3781 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3841 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3901 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3961 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4021 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4081 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4141 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4201 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4261 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4321 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4381 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4441 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4501 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4561 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4621 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4681 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4741 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4801 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4861 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4921 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4981 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5041 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5101 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5161 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5221 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5281 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5341 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5401 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5461 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5521 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5581 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5641 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5701 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5761 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5821 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5881 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5941 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6001 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6061 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6121 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6181 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6241 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6301 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6361 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6421 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6481 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6541 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6601 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6661 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6721 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6781 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6841 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6901 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     6961 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7021 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7081 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     7141 gtt
//
