LOCUS       sequence-174775-        8173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of SZT2 fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8173)
  AUTHORS   Wolfson RL, Chantranupong L, Wyant GA, Gu X, Orozco JM, Shen K, 
            Condon KJ, Petri S, Kedir J, Scaria SM, Abu-Remaileh M, Frankel WN, 
            Sabatini DM
  TITLE     KICSTOR recruits GATOR1 to the lysosome and is necessary for 
            nutrients to regulate mTORC1.
  JOURNAL   Nature. 2017 Feb 15. doi: 10.1038/nature21423.
  PUBMED    28199306
REFERENCE   2  (bases 1 to 8173)
  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: "Nature. 
            2017 Feb 15. doi: 10.1038/nature21423."
COMMENT     Sequence Label: pRK5-HA-SZT2 2241-end
FEATURES             Location/Qualifiers
     source          1..8173
                     /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     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      950..959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             962..988
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      4460..4469
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4473..4607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4510..4529)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4564..4583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4676..5033
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4676..4867
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4699..4719)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4884..5019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5053..5070)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5053..5069)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5062..5084)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5282..5737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5369..5388)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5579..5600
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5754..5773)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5873..5895
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5933..5951)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6019..6123
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6124..6192,6193..6984)
                     /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     complement(6342..6361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7155..7743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7644..7663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7897..7914
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8031..8052
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8067..8072,8073..8090,8091..8097)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8105..8121
                     /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     8110..8132
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8129..8145
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8129..8145
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
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 agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgattgaatt cccgaaccga cagtcggtct cttcaccaag gccattcgcg ccaccatggg
      961 ctacccctat gatgtgcctg actacgcaga tctcaatggt ggtggtggtg ggtcgacgac
     1021 tctagccttt gtggatgaag gaggggcccc cttgtcactg gcgttgtggc ccccctcctc
     1081 tccggggccc ccagacccac tgcgagagga ggaatttgag caactgaccc aggtcatccg
     1141 ctgcccggtt gttgtggaca gttcttcagc tcagaatggg gccccacggc ttcgattgga
     1201 tgtgtgggaa aaggggaaca ttagtattgt gcagctggag gagaaactcc gaggagcagc
     1261 tcgccaggcc ctggccgatg ccatcatcga gcttcagctg ctgccagctt cactatgtac
     1321 agaggacaca cccacaggaa gtctcaggaa cggatcgttg gaaactaaga gctctgcagg
     1381 ccgagctagc acctttcccc ctgcccctgt ccctggggag cctgtgactc cacccagcaa
     1441 agcgggccgg cgtagcttct gggatatgct gagtaaaaca gaatgtgggg atttgggttc
     1501 ccccaaaaca actgatgaca ttgtcctgga tcggccagaa gacactcggg gccggaggcg
     1561 tcacaaaacc gagagtgttc ggactcctgg tggagctgag cgggcgccag gctcagattc
     1621 tggagcccag agacaaaagc gccggacaac acagctagaa gagggtgagg tggggaccct
     1681 tcatcctgtg tttgcccgtg ttgctcagcg ctggatggag tttatggttc agattggttg
     1741 tgcctcagtg tccagaagct ctgcccacat ggtgtcccgg ttcctccttc catccatcct
     1801 gtctgagttc accgcactgg tcacctcaat ggctggagac accagtgtcc gcatctttga
     1861 gcagcatttg ggttcagagc cagagatctt cggcccttgt tcccctgggc aactgggccc
     1921 ctctccccgc cctgcagctg agcggcatct gctgcttctg ggaaggaact tcttgcagtg
     1981 gaggagacca acacagcagg ctgccaaagc catgcagcgc ttcgagccag gaggtgatgg
     2041 gagctcaggg cgaaatgctc cccggcagag gctcttgcta ctagaggttg tggacaagaa
     2101 gctacagctg ctgacctaca actgggctcc agacctgggg gcagcattgg gccgagcgct
     2161 ggttcgcctg gtgcagtggc agaatgcacg agcccatctc atcttctgcc tactcagcca
     2221 gaagcttggc ctcttccatc attatggcca gttggacttc cccgtgcgag atgaaaagga
     2281 gccaaaccca ttcctgctgc cgaccatgga agtggagacc ctcatccgga gtgcaagtcc
     2341 cccgctgagc cgtgagcagg gccgactgag tgggtcctct cgtggtgggg gtcctcttcc
     2401 cctggacaca ttcccctttg acgaggccct aagggatatc acggctgccc gccccagctc
     2461 cgtacttggt cctgtgccca gacctcctga tcctgtcacc taccatggac aacagttcct
     2521 agagatcaag atggcagagc gcagagagct ggagcgccag atgaagatgg aaaacctgtt
     2581 tgtaacctgg cagcagcgtt ctaccccagc caccatgccc atcagtgctg gagagctgga
     2641 gaccctgaag cagtcatccc gcctggtgca ttactgtgca acagccatgc tcttcgaccc
     2701 agctgcctgg ctgcatgggc ccccagagac ctctggaccc cctgacgggc agcggcgcca
     2761 tcgccctgag tcagggtctg ggagccgaga ggcccccaca agctgtgaat ccttggatgt
     2821 gtcgcccccg ggagcccgtg aggagccttg gctgaaggag ctgagcttgg ctttcctgca
     2881 gcaatatgtg cagtatctgc agagcatagg ttttgtgctg gtaccactgc ggcccccctc
     2941 acccgcccgc agcaccagcc ggccacgggc catggctatc cttggaacag agggtcgagg
     3001 ctccttctcc tgccctaaaa ccaagactga tgggagcccc aagagcacta gctctccggt
     3061 aaccacctac cacctgcagc gggcactgcc tgggggcatc atcctcatgg aactggcatt
     3121 ccagggctgt tacttctgtg tcaaacagtt tgccctggaa tgttcccgaa tcccaatggg
     3181 gcaggctgtc aactcacagc tgtccatgct gttcacagag gagtgtgaca aggtgcggga
     3241 cctgatgcac gtgcactcgt tcagctatga cttccatctg cgcctcgtgc atcagcacgt
     3301 gctaggtgcc catctggtgc tgcggcacgg ctaccacctc accacctttc tgcgacactt
     3361 cctggcccac caccctgacg gaccccactt tggccgcaat cacatttacc aagggacatt
     3421 ggagctcccc acaccactca ttgctgccca ccagctatac aactacgtgg ctgatcacgc
     3481 cagctcttac cacatgaagc cattgcgaat ggcccggcca gggggcccag aacacaacga
     3541 gtatgccctg gtgtcggcat ggcacagttc tggctcctac ctggactctg agggacttcg
     3601 acaccaggat gactttgatg tgtctctgct tgtctgtcac tgtgctgcac cctttgagga
     3661 gcaaggagag gctgagcggc acgttctgcg gctacagttc ttcgtggtgc tcaccagcca
     3721 gcgagagctc ttccccaggc tcactgctga catgcgccgc ttccggaagc cacccagact
     3781 gccccctgag ccagaggctc ctgggagttc agctggcagc cctggggagg cctcagggct
     3841 tattctagcg cctggaccgg ctcctctgtt cccaccactg gctgcagagg tgggcatggc
     3901 acgagcacgg ctggctcagc tggtgcggct ggctggaggg cactgccgtc gggacaccct
     3961 ttggaagcgc ctcttcttgc tggagccacc ggggcctgat cgactgcggc taggggggcg
     4021 cctggccctg gcagagctgg aggaactcct agaagcagtc catgccaaat ccattgggga
     4081 catcgacccc cagctggact gcttcctatc catgacggtc tcctggtacc agagcctgat
     4141 caaagttctc ctaagccgct tcccccagag ctgtcgccat ttccaaagcc cagacttggg
     4201 aactcagtac ctggttgtgc tgaatcagaa gttcactgac tgctttgtgc tagtgtttct
     4261 ggactcccac ttaggaaaga cgtctctgac agtggttttc cgagagccct tcccagtaca
     4321 gccccaggac agcgagagcc cccctgccca actggtctcc acctaccacc acctggagtc
     4381 tgtcatcaac acagcctgtt tcaccctctg gacccgcctc ctctgagcgg ccgctaagta
     4441 agtaaggatc cccagcttgg ccgccatggc ccaacttgtt tattgcagct tataatggtt
     4501 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4561 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcggg aattaattcg
     4621 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
     4681 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4741 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4801 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4861 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4921 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4981 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctgt
     5041 taacagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5101 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5161 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     5221 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     5281 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5341 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5401 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5461 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     5521 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5581 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
     5641 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5701 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     5761 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     5821 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5881 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5941 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     6001 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6061 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6121 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6181 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6241 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6301 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6361 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6421 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6481 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6541 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6601 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6661 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6721 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6781 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6841 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6901 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6961 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7021 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7081 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7141 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7201 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7261 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7321 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7381 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7441 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7501 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7561 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7621 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7681 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7741 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7801 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7861 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7921 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7981 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     8041 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     8101 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattaa
     8161 ttcgagctcg ccc
//
