LOCUS       sequence-400985-        7438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FIP200 NTD-MBP mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7438)
  AUTHORS   Chen M, Ren X, Nguyen TN, Khuu G, Cook ASI, Lazarou M, Hurley JH
  TITLE     Structure and activation of the human autophagy-initiating 
            ULK1C:PI3KC3-C1 supercomplex
  JOURNAL   bioRxiv 2023.06.01.543278
REFERENCE   2  (bases 1 to 7438)
  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: "bioRxiv 
            2023.06.01.543278"
COMMENT     Sequence Label: pCAG-FIP200  (1-640)-GST
FEATURES             Location/Qualifiers
     source          1..7438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        103..482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        484..759
                     /label=chicken beta-actin promoter
     intron          760..1776
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1700..1722
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1784..1803
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3782..3802
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3806..4459
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4417..4439
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     primer_bind     complement(4558..4577)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4623..4678
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4677..4696)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5039..5055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5039..5055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5052..5074)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5063..5079
                     /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(5087..5093,5094..5111,5112..5117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5132..5153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5212..5407
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5258..5393
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5413..5547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5450..5469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5504..5523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5615..5632)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5786..6374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5866..5885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6545..7336,7337..7405))
                     /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     7168..7187
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7406..7438,1..72))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
       61 aggggttccg cgcacatttc cccgaaaagt gccacctggg tcgacattga ttattgacta
      121 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      181 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      241 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      301 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      361 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      421 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      481 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
      541 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg
      601 gggsgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
      661 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      721 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      781 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      841 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      901 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
      961 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
     1021 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
     1081 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
     1141 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
     1201 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
     1261 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
     1321 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
     1381 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     1441 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     1501 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     1561 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1621 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     1681 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     1741 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     1801 gtgctgtctc atcattttgg caaagaattc ctcgatcgac ggtatcgatg ccaccatgaa
     1861 gctctatgtg tttctcgtga ataccggcac taccctgacc tttgataccg aactgaccgt
     1921 ccagaccgtg gctgacctga agcatgctat ccagtctaaa tacaagatcg caattcagca
     1981 ccaggtgctg gtggtcaacg gcggagagtg catggccgct gatcgcaggg tctgtaccta
     2041 ttcagccggc accgacacta acccaatctt cctctttaat aaggagatga ttctgtgcga
     2101 tagaccccct gctatcccca agaccacttt tagcactgag aacgacatgg aaatcaaagt
     2161 ggaggaatcc ctgatgatgc cagccgtgtt ccacaccgtc gcttctcgca ctcagctggc
     2221 actcgagatg tacgaagtgg ccaagaaact gtgctctttc tgtgagggtc tcgtccacga
     2281 tgaacatctg cagcaccagg gctgggcagc cattatggct aacctggagg actgttcaaa
     2341 tagttatcag aaactgctct tcaagtttga gtctatctac tcaaactatc tgcagagcat
     2401 cgaagacatt aaactgaagc tcacacacct gggaaccgca gtgagcgtca tggccaagat
     2461 ccctctgctc gagtgcctga ctaggcattc ctacagagaa tgtctgggta ggctcgatag
     2521 tctgccagaa cacgaggaca gcgagaaagc cgaaatgaag cgctctacag agctggtgct
     2581 ctcacctgac atgccaagga caaccaacga aagtctgctc accagcttcc caaagtccgt
     2641 ggagcatgtc tcccccgaca ctgccgatgc tgaatctgga aaggagatta gagaaagttg
     2701 ccagagcacc gtgcaccagc aggacgagac tacaattgac actaaggacg gcgatctgcc
     2761 cttctttaac gtcagtctgc tcgattggat caacgtccag gacagaccta atgatgtgga
     2821 gtccctggtc cggaagtgct ttgatagcat gtcccgtctg gacccacgta tcattcgtcc
     2881 tttcatcgca gagtgtcgtc agaccattgc caagctggac aaccagaata tgaaagctat
     2941 taagggtctg gaagatcggc tctacgccct ggaccagatg atcgctagct gtggacgcct
     3001 ggtgaacgag cagaaggaac tcgctcaggg tttcctggca aaccagaaaa gagccgagaa
     3061 tctcaaggat gcatcagtgc tgcctgacct gtgcctcagt catgccaatc agctgatgat
     3121 tatgctccag aaccaccgca agctgctcga catcaaacag aagtgtacca ctgctaaaca
     3181 ggagctggca aacaatctcc atgtgaggct gaagtggtgc tgtttcgtca tgctgcacgc
     3241 agaccaggat ggcgagaagc tccaggccct gctcaggctg gtgattgagc tgctcgaaag
     3301 agtgaagatc gtcgaggctc tgtccaccgt cccacagatg tactgcctgg cagtggtcga
     3361 ggtggtcaga cgtaaaatgt ttatcaagca ctacagagaa tgggcaggtg ccctcgtgaa
     3421 agatggcaag cgtctgtatg aagccgagaa atctaagcgt gagtcattcg gcaaactgtt
     3481 tcggaagagc ttcctccgta accggctgtt ccgcggactc gacagttggc cacccagctt
     3541 ttgcacccag aaacctagga agttcgactg tgagctgcca gatatctccc tcaaggacct
     3601 gcagtttctc cagtccttct gcccatctga ggtgcagccc tttctgaggg tccctctgct
     3661 ctgtgacttc gaacctctgc accagcatgt gctggccctc cacaacctcg tcaaggctgc
     3721 acagagtctg gatgagatga gccagacaat caccgacctg ctcagcgaac agaagggtac
     3781 cgaaaacctg tacttccaat ccggtatgtc ccctatacta ggttattgga aaattaaggg
     3841 ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt
     3901 gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt
     3961 tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat
     4021 acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat
     4081 ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag
     4141 taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat
     4201 gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga
     4261 cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc
     4321 gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta
     4381 cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg
     4441 tggcgaccat cctccaaaaa ttgactagct cgaggaattc actcctcagg tgcaggctgc
     4501 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     4561 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     4621 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     4681 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     4741 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
     4801 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     4861 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     4921 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     4981 tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt
     5041 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5101 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5161 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa
     5221 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5281 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5341 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5401 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5461 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5521 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga
     5581 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5641 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5701 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5761 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5821 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5881 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5941 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6001 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6061 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6121 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6181 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6241 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6301 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6361 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6421 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6481 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6541 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6601 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6661 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6721 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6781 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6841 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6901 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6961 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7021 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7081 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7141 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7201 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7261 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7321 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7381 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcattta
//
