LOCUS       sequence-244473-        7903 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 7903)
  AUTHORS   Hosokawa N, Hara T, Kaizuka T, Kishi C, Takamura A, Miura Y, Iemura 
            S, Natsume T, Takehana K, Yamada N, Guan JL, Oshiro N, Mizushima N
  TITLE     Nutrient-dependent mTORC1 association with the ULK1-Atg13-FIP200 
            complex required for autophagy.
  JOURNAL   Mol Biol Cell. 2009 Apr . 20(7):1981-91.
  PUBMED    19211835
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: "Mol Biol 
            Cell. 2009 Apr . 20(7):1981-91."
COMMENT     Sequence Label: pME18s-HA-hFIP200
FEATURES             Location/Qualifiers
     source          1..7903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..330,331..345,346..614)
                     /label=SR-alpha promoter
                     /note="hybrid promoter consisting of the SV40 early 
                     promoter plus part of the long terminal repeat of human 
                     T-lymphotrophic virus 1"
     rep_origin      181..316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             346..614
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          644..740
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             793..819
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             823..849
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             853..879
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3567..3578
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3810..3821
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5690..5706)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5690..5706)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    5791..5925
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5828..5847)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5882..5901
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5964..5981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6135..6723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6215..6234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6894..7685,7686..7754))
                     /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     7517..7536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7755..7859)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(7899..7903,1..16))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
       61 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      241 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
      301 ggcttttttg gaggcctagg cttttgcaaa aagctcctcg atcgaggggc tcgcatctct
      361 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
      421 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
      481 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
      541 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
      601 cgccgttaca gatccaagct ctgaaaaacc agaaagttaa ctggtaagtt tagtcttttt
      661 gtcttttatt tcaggtcccg gatccggtgg tggtgcaaat caaagaactg ctcctcagtg
      721 gatgttgcct ttacttctag gcctgtacgg aagtgttact tctgctctaa aagctgcgga
      781 attgccatgg cataccctta tgacgtacca gattacgcgg gatatcccta cgatgtcccc
      841 gattacgcgc gttatccata tgacgtcccg gactatgccg aattcatgaa gttatatgta
      901 tttctggtta acactggaac tactctaaca tttgacactg aacttacagt gcaaactgtg
      961 gcagacctta agcatgccat tcaaagcaaa tacaagattg ctattcaaca ccaggtgctg
     1021 gtggtcaatg gaggagaatg catggctgca gatcgaagag tgtgtaccta cagtgctggg
     1081 acggatacaa atccaatttt tctttttaac aaagaaatga tcttatgtga tcgtccacct
     1141 gctattccta aaactacctt ttcgacagaa aatgacatgg aaataaaagt tgaagaatct
     1201 cttatgatgc ctgcagtttt tcatactgtt gcttcaagga cacagcttgc attggaaatg
     1261 tatgaagttg ccaagaaact ttgttctttt tgtgaaggtc ttgtacatga tgaacatctt
     1321 caacaccaag gctgggctgc aatcatggcc aacctggagg actgttcaaa ttcataccaa
     1381 aagctacttt tcaagtttga aagtatttat tcaaattatc tgcagtccat agaagacatc
     1441 aagttaaaac ttactcattt aggaactgca gtttcagtaa tggccaagat tccactgttg
     1501 gagtgcctaa ccagacatag ttacagagaa tgtttgggaa gactggattc tttacctgaa
     1561 catgaagact cagaaaaagc tgagacgaaa agatccactg aactggtgct ctctcctgat
     1621 atgcctagaa caactaacga atctttgtta acctcatttc ccaagtcagt ggaacatgtg
     1681 tccccagata ccgcagatgc tgaaagtggc aaagaaatta gggaatcttg tcaaagtact
     1741 gttcatcagc aagatgaaac tacgattgac actaaagatg gtgatctgcc cttttttaat
     1801 gtctctttgt tagactggat aaatgttcaa gatagaccta atgatgtgga atctttggtc
     1861 aggaagtgct ttgattctat gagcaggctt gatccaagga ttattcgacc atttatagca
     1921 gaatgccgtc aaactattgc caaacttgat aatcagaata tgaaagccat taaaggactt
     1981 gaagatcggc tctacgccct ggaccagatg attgctagct gtggccgact ggtgaatgaa
     2041 cagaaagagc ttgctcaggg atttttagct aatcagaaga gagctgaaaa cttaaaggat
     2101 gcatctgtat tacctgattt atgcctgagt cacgcaaatc agttgatgat tatgttgcaa
     2161 aatcatagaa aactgttaga tattaagcag aagtgtacca ctgccaaaca agaactagca
     2221 aataacctac atgtcagact gaagtggtgt tgctttgtaa tgcttcatgc tgatcaagat
     2281 ggagagaagt tacaagcttt gctccgcctc gtaatagagc tgttagaaag agtcaaaatt
     2341 gttgaagctc ttagtacagt tcctcagatg tactgcttag ctgttgttga ggttgtaaga
     2401 agaaaaatgt tcataaaaca ctacagggag tgggctggtg ctttagtcaa agatggaaag
     2461 agattatatg aagcagaaaa atcaaaaagg gaatcctttg ggaaattatt taggaagtct
     2521 tttttaagaa atcgtctgtt taggggactg gactcctggc ccccttcctt ttgtactcaa
     2581 aagcctcgaa agtttgactg tgaacttcca gatatttcat taaaagattt acagtttctg
     2641 caatcatttt gtccttcgga agttcagcca ttcctcaggg ttcccttact ttgtgacttt
     2701 gaacctctac accagcatgt acttgctcta cataatttgg taaaagcagc acaaagtttg
     2761 gatgaaatgt cacagaccat tacagatcta ctgagtgaac aaaaggcatc tgtgagtcag
     2821 acatccccac agtctgcttc ttcaccaagg atggaaagta cagcaggaat tacaactact
     2881 acctcaccga gaactcctcc accactgact gttcaggatc ccttatgtcc tgcagtttgt
     2941 cccttagaag aattatctcc agatagtatt gatgcacata cgtttgattt tgaaactatt
     3001 ccccatccaa acatagaaca gactattcac caagtttctt tagacttgga ttcattagca
     3061 gaaagtcctg aatcagattt tatgtctgct gtgaatgagt ttgtaataga agaaaatttg
     3121 tcgtctccta atcctataag tgatccacaa agcccagaaa tgatggtgga atcactttat
     3181 tcatcagtta tcaatgcgat agacagtaga cgaatgcagg atacaaatgt atgtggtaag
     3241 gaggattttg gagatcatac ttctctgaat gtccagttgg aaagatgtag agttgttgcc
     3301 caagactctc acttcagtat acaaaccatt aaggaagacc tttgccactt tagaacattt
     3361 gtacaaaaag aacagtgtga cttctcaaat tcattaaaat gtacagcagt agaaataaga
     3421 aacattattg aaaaagtaaa atgttctctg gaaataacac taaaagaaaa acatcaaaaa
     3481 gaactactgt ctttaaaaaa tgaatatgaa ggtaaacttg acggactaat aaaggaaact
     3541 gaagagaatg aaaacaaaat taaaaaattg aagggagagt tagtatgcct tgaggaggtt
     3601 ttacaaaata aagataatga atttgctttg gttaaacatg aaaaagaagc tgtaatctgc
     3661 ctgcagaatg aaaaggatca gaagttgtta gagatggaaa atataatgca ctctcaaaat
     3721 tgtgaaatta aagaactgaa gcagtcacga gaaatagtgt tagaagactt aaaaaagctc
     3781 catgttgaaa atgatgagaa gttacagtta ttgagggcag aacttcagtc cttggagcaa
     3841 agtcatctaa aggaattaga ggacacactt caggttaggc acatacaaga gtttgagaag
     3901 gttatgacag accacagagt ttctttggag gaattaaaaa aggaaaacca acaaataatt
     3961 aatcaaatac aagaatctca tgctgaaatt atccaggaaa aagaaaaaca gttacaggaa
     4021 ttaaaactca aggtttctga tttgtcagac acgagatgca agttagaggt tgaacttgcg
     4081 ttgaaggaag cagaaactga tgaaataaaa attttgctgg aagaaagcag agcccagcag
     4141 aaggagacct tgaaatctct tcttgaacaa gagacagaaa atttgagaac agaaattagt
     4201 aaactcaacc aaaagattca ggataataat gaaaattatc aggtgggctt agcagagcta
     4261 agaactttaa tgacaattga aaaagatcag tgtatttccg agttaattag tagacatgaa
     4321 gaagaatcta atatacttaa agctgaatta aacaaagtaa catctttgca taaccaagca
     4381 tttgaaatag aaaaaaacct aaaagaacaa ataattgaac tgcagagtaa attggattca
     4441 gaattgagtg ctcttgaaag acaaaaagat gaaaaaatta cccaacaaga agagaaatac
     4501 gaagctatta tccagaacct tgagaaagac agacaaaaat tggtcagcag ccaggagcaa
     4561 gacagagaac agttaattca gaagcttaat tgtgaaaaag atgaagctat tcagactgcc
     4621 ctaaaagaat ttaaattgga gagagaagtt gttgagaaag agttattaga aaaagttaaa
     4681 catcttgaga atcaaatagc aaaaagtcct gccattgact ctaccagagg agattcttca
     4741 agcttagttg ctgaacttca agaaaagctt caggaagaaa aagctaagtt tctagaacaa
     4801 cttgaagagc aagaaaaaag aaagaatgaa gaaatgcaaa atgttcgaac atctttgatt
     4861 gcggaacaac agaccaattt taacactgtt ttaacaagag agaaaatgag aaaagaaaac
     4921 ataataaatg atcttagtga taagttgaaa agtacaatgc agcaacaaga acgggataaa
     4981 gatttgatag agtcactttc tgaagatcga gctcgtttgc ttgaggaaaa gaaaaagctt
     5041 gaagaagaag tcagtaagtt gcgcagtagc agttttgttc cttcaccata tgtagctaca
     5101 gccccagaac tttatggagc ttgtgcacct gaactcccag gtgaatcaga tagatccgct
     5161 gtggaaacag cagatgaagg aagagtggat tcagcaatgg agacaagcat gatgtctgta
     5221 caagaaaata ttcatatgtt gtctgaagaa aaacagcgga taatgctgtt agaacgaaca
     5281 ttgcaattga aagaagaaga aaataaacgg ttaaatcaaa gactgatgtc tcagagcatg
     5341 tcttcagtat cttcaaggca ttctgaaaag atagctatta gagattttca ggtgggagat
     5401 ttggtactca tcatcctaga cgaacgccat gacaattatg tgttatttac tgttagtcct
     5461 actttatatt ttctacattc agagtctcta cctgccctgg atctcaaacc aggtgagggt
     5521 gcttcaggtg catctagaag accctgggta ctcggaaaag taatggaaaa agaatactgt
     5581 caagccaaaa aggcacaaaa cagatttaaa gttcctttgg ggacaaagtt ttacagagtg
     5641 aaagccgtat catggaataa gaaagtataa gaattcctgc agcccggggg atccactagt
     5701 tctagagcgg ccgctagact agtctagaga aaaaacctcc cacacctccc cctgaacctg
     5761 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     5821 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5881 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccccgg gtaccgagct
     5941 cgaattaatt cctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6001 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6061 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6121 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6181 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6241 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6301 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6361 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6421 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6481 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6541 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6601 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6661 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6721 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6781 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6841 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6901 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6961 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7021 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7081 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7141 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7201 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7261 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7321 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7381 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7441 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7501 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7561 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7621 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7681 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7741 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7801 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7861 acatttcccc gaaaagtgcc acctgcaagc ttggctgtgg aat
//
