LOCUS       sequence-174765-        7828 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 7828)
  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 7828)
  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 1-1021
FEATURES             Location/Qualifiers
     source          1..7828
                     /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      4115..4124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4128..4262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4165..4184)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4219..4238
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4331..4688
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4331..4522
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4354..4374)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4539..4674
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4708..4725)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4708..4724)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4717..4739)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4937..5392
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5024..5043)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5234..5255
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5409..5428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5528..5550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5588..5606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5674..5778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5779..5847,5848..6639)
                     /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(5997..6016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6810..7398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7299..7318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7552..7569
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7686..7707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7722..7727,7728..7745,7746..7752)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7760..7776
                     /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     7765..7787
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7784..7800
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7784..7800
                     /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 ggtcgacgat
     1021 ggcctcggag cgcccggagc cggaggtgga agaagctggg caggtgttcc tgttaatgaa
     1081 aaaggattat cgaatctccc gaaatgttcg cctggcttgg ttcctcagtc atctgcacca
     1141 aactgtgcag gccacacccc aggagatgct gcttcagtct gaacaggaat tggaagtcct
     1201 cagtgtcctg ccccctgggt ggcagccaga tgaaccagtg gtcccaaggc cattcctcct
     1261 ggtaccttcc acccgggtca ccttcctggc ttggcagtat cggtttgtca ttgagttgga
     1321 ccttagccca tctactggca ttgtggatga ttccacaggg gagatcttgt ttgatgaagt
     1381 tttccatgcc ctgtcccgct gcttaggcgg gctgcttcgg cccttccgag tgcctggatc
     1441 ttgcatcgac ttccagcctg agatctatgt aactatccag gcctactcct ccatcattgg
     1501 actgcagtcc caccaggtgc tggtacaggg ctgcctcttg gacccttccc agcgggaggt
     1561 gttcctgcag cagatatatg agcagctctg cctctttgag gataaggtgg ccaccatgct
     1621 gcagcagcag tacgatcccc agagccaggc agaagaccag tccccagact caggggacct
     1681 actgggccgg aaggtaggcg tctccatggt gacagctgat cttgggctgg tcagtatgat
     1741 tcgtcagggc atcttggcac tgcagttact accctcgaac tctagtgcag ggattatcgt
     1801 gatcacggat ggggtgacca gtgtacctga tgttgctgtc tgtgagacac tgctgaacca
     1861 gcttcgcagt ggcactgtgg cttgttcctt tgtccaggtg ggaggagttt actcttatga
     1921 ctgcagtttt ggccatgtgc ccaatgtgga attaatgaag ttcatcgcaa tggcaacatt
     1981 tgggtcctac ctgtccactt gtcctgagcc ggagccaggc aacctgggtc tgactgtcta
     2041 ccaccgggca tttctcctct attccttcct gcgcagtggg gaagcactga accctgaata
     2101 ttactgcggc tctcagcacc gcctatttaa tgagcacctg gtctctgcaa gcagcaaccc
     2161 tgccctggcc ttgcgccgga agaagcacac tgagaaggag gtgccagccg acttggtcag
     2221 cactgtgtcc gtacggcttc gagagggcta cagtgtccga gaggtcacac tggccaaagg
     2281 agggtcccaa ttggaggtaa agctggtgct gctgtggaaa cacaacatgc gcattgagta
     2341 tgtggctatg gcaccctggc ccctggagcc tgagggccct cgagtaacac gggtggaagt
     2401 gacgatggaa ggcggctacg acattttgca tgatgtgtcc tgtgcactaa ggcagcccat
     2461 tcgttcattg tatcgtaccc atgttatccg gcgtttctgg aacacgctgc agagcatcaa
     2521 ccagacagac cagatgcttg cccaccttca gtccttctcc tcagtgcctg agcatttcac
     2581 gcttcctgac agcaccaaga gcggagtgcc actcttctac atccctccag gctccaccac
     2641 cccggtgctc tccctccagc ccagtggttc tgactcatcc catgcccagt ttgctgccta
     2701 ctggaagcca gtgctgtcca tggatgcaaa ttcctggcag cgatggctgc acatgcatcg
     2761 cctggtgcta atcctggagc atgacacacc aatccccaag cacttgcaca ccccgggcag
     2821 caatgggcgc tacagcacta tccagtgcag gatctcccac tcctccctga cctctctgct
     2881 gcgggactgg agcagcttcg tactagtcga gggctattct tatgttaagc tgctctccag
     2941 tgccccagac cagcccccca attccttcta catggtccgt atcatttcca aggccccatg
     3001 catggttctt cgcctgggtt ttcccattgg cacaccagca ccggcccggc acaagattgt
     3061 gtcaggcttg agggaagaga tcctgcggct gcgtttcccc caccgggtac aaagcaagga
     3121 gccaacgccc aaggtgaaac gaaaagggct agggggtgct ggtgggggca gctctccctc
     3181 caagtcaccc cccgtgctgg ggccacagca ggccctgtct gaccggccct gccttgtggt
     3241 cctgcataag ccactggaca aactgctcat caggtatgag aagctgccct tggactaccg
     3301 ggcacccttc ttgctgacat tggagccacc aggtccactg cccttggtgt caggccgctc
     3361 agcctcttct agcctggcgt cactgtcccg ctacctctac catcagcgct ggctttggag
     3421 tgtcccgtca ggactggccc ctgcgctgcc tctcagtgcc attgcccagc tcctctccat
     3481 cctcactgaa gtccgacttt ctgaaggatt ccacttcgcc tgcagtgggg aaggaatcat
     3541 caacatggtt ctggagcttc caattcagaa tgaaccacca gggcaggctg cagctgaaga
     3601 gaagcacacc tgtgttgtcc agtacatcct cttcccccca cactctacct ccaccaaaga
     3661 cagcttctcg acagatgatg acaatgatgt ggaagtggag gccctggagg gagactcaga
     3721 gctcaatctg gtcactgagg tgtgggtgga gccacagtat gggcgagtgg gacctggccc
     3781 tggaatctgg aagcacctcc aggacctgac gtattctgag atcccgcaag ctctccaccc
     3841 acgggatgct gcctgcatag gctccatgct gagctttgaa tacctgatac agctgtgtca
     3901 gagcaaggaa tggggtcctc tgcccccaga gccgagggtc tctgatggat tggatcaggg
     3961 aggagacacc tgcgtccatg agatcccttt ccattttgac ctaatgggat tgctgccaca
     4021 gtgccagcag ctccagatgt tcttcctctt gcttgccaga ggtgccgagg ggccactgtg
     4081 agcggccgct aagtaagtaa ggatccccag cttggccgcc atggcccaac ttgtttattg
     4141 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4201 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     4261 tcgggaatta attcggcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     4321 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4381 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4441 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4501 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4561 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4621 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4681 tttgcaaaaa gctgttaaca gcttggcact ggccgtcgtt ttacaacgtc gtgactggga
     4741 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     4801 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4861 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4921 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4981 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5041 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5101 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     5161 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5221 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     5281 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5341 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     5401 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     5461 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5521 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5581 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5641 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5701 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5761 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5821 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5881 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5941 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6001 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6061 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6121 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6181 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6241 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6301 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6361 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6421 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6481 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6541 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6601 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6661 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6721 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6781 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6841 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6901 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6961 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7021 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7081 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7141 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7201 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7261 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7321 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7381 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7441 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7501 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7561 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7621 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7681 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7741 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     7801 atgattacga attaattcga gctcgccc
//
