LOCUS       sequence-129746-        7260 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7260)
  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 7260)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-HA
FEATURES             Location/Qualifiers
     source          1..7260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2706..2732
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2758..2799
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2809..2826
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2849..2866)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2855..3079
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3125..3553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3212..3231)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3422..3443
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3562..3582)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3567..3896
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3747..3882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3809..3828
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3963..4757
                     /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(4017..4036)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4627..4646
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4933..5054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4970..4989)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5024..5043
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5103..5119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5103..5119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5116..5138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5127..5143
                     /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(5151..5157,5158..5175,5176..5181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5196..5217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5334..5351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5505..6093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5585..5604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6264..7055,7056..7124))
                     /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     6887..6906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7125..7229)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattgcc cttgccacca
      961 tgacagcccc ggcgggtccg cgcggctcag agaccgagcg gcttctgacc cccaaccccg
     1021 ggtatgggac ccaggcgggg ccttcaccgg cccctccgac acccccagaa gaggaagacc
     1081 ttcgccgtcg tctcaaatac tttttcatga gtccctgcga caagtttcga gccaagggcc
     1141 gcaagccctg caagctgatg ctgcaagtgg tcaagatcct ggtggtcacg gtgcagctca
     1201 tcctgtttgg gctcagtaat cagctggctg tgacattccg ggaagagaac accatcgcct
     1261 tccgacacct cttcctgctg ggctactcgg acggagcgga tgacaccttc gcagcctaca
     1321 cgcgggagca gctgtaccag gccatcttcc atgctgtgga ccagtacctg gcgttgcctg
     1381 acgtgtcact gggccggtat gcgtatgtcc gtggtggggg tgacccttgg accaatggct
     1441 cagggcttgc tctctgccag cggtactacc accgaggcca cgtggacccg gccaacgaca
     1501 catttgacat tgatccgatg gtggttactg actgcatcca ggtggatccc cccgagcggc
     1561 cccctccgcc ccccagcgac gatctcaccc tcttggaaag cagctccagt tacaagaacc
     1621 tcacgctcaa attccacaag ctggtcaatg tcaccatcca cttccggctg aagaccatta
     1681 acctccagag cctcatcaat aatgagatcc cggactgcta taccttcagc gtcctgatca
     1741 cgtttgacaa caaagcacac agtgggcgga tccccatcag cctggagacc caggcccaca
     1801 tccaggagtg taagcacccc agtgtcttcc agcacggaga caacagcttc cggctcctgt
     1861 ttgacgtggt ggtcatcctc acctgctccc tgtccttcct cctctgcgcc cgctcactcc
     1921 ttcgaggctt cctgctgcag aacgagtttg tggggttcat gtggcggcag cggggacggg
     1981 tcatcagcct gtgggagcgg ctggaatttg tcaatggctg gtacatcctg ctcgtcacca
     2041 gcgatgtgct caccatctcg ggcaccatca tgaagatcgg catcgaggcc aagaacttgg
     2101 cgagctacga cgtctgcagc atcctcctgg gcacctcgac gctgctggtg tgggtgggcg
     2161 tgatccgcta cctgaccttc ttccacaact acaatatcct catcgccaca ctgcgggtgg
     2221 ccctgcccag cgtcatgcgc ttctgctgct gcgtggctgt catctacctg ggctactgct
     2281 tctgtggctg gatcgtgctg gggccctatc atgtgaagtt ccgctcactc tccatggtgt
     2341 ctgagtgcct gttctcgctc atcaatgggg acgacatgtt tgtgacgttc gccgccatgc
     2401 aggcgcagca gggccgcagc agcctggtgt ggctcttctc ccagctctac ctttactcct
     2461 tcatcagcct cttcatctac atggtgctca gcctcttcat cgcgctcatc accggcgcct
     2521 acgacaccat caagcatccc ggcggcgcag gcgcagagga gagcgagctg caggcctaca
     2581 tcgcacagtg ccaggacagc cccacctccg gcaagttccg ccgcgggagc ggctcggcct
     2641 gcagccttct ctgctgctgc ggaagggacc cctcggagga gcattcgctg ctggtgaatc
     2701 tcgagtaccc atatgatgtt cctgattacg cgtaatctag agggcccgcg gttcgaaggt
     2761 aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat
     2821 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2881 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2941 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3001 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3061 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3121 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3181 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3241 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3301 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3361 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3421 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3481 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3541 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3721 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3841 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3901 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3961 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4021 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4081 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4141 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4201 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4261 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4321 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4381 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4441 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4501 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4561 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4621 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4681 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4741 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     4801 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4861 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4921 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4981 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5041 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5101 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5161 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5221 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5281 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5341 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5401 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5461 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5521 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5581 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5641 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5701 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5761 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5821 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5881 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5941 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6001 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6061 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6121 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7141 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
