LOCUS       sequence-363234-        7918 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 7918)
  AUTHORS   Jung CH, Jun CB, Ro SH, Kim YM, Otto NM, Cao J, Kundu M, Kim DH
  TITLE     ULK-Atg13-FIP200 complexes mediate mTOR signaling to the autophagy 
            machinery.
  JOURNAL   Mol Biol Cell. 2009 Apr;20(7):1992-2003. Epub 2009 Feb 18.
  PUBMED    19225151
REFERENCE   2  (bases 1 to 7918)
  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."; date: "2009-04"; volume: "20(7)"; pages: "1992-2003. Epub 
            2009 Feb 18"
COMMENT     Sequence Label: HA-hULK1
FEATURES             Location/Qualifiers
     source          1..7918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2113..2139)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2142..2151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2236..2255)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2268..2286)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2269..2286)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2496..2520)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2517..2720)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2546..2566)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2721..3100
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3129..3145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3129..3145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3142..3164)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3153..3169
                     /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(3177..3183,3184..3201,3202..3207))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3222..3243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3360..3377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3531..4119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3611..3630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4290..5081,5082..5150))
                     /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     4913..4932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5151..5255)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5323..5341
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5379..5401)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5501..5520
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5537..5992)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5674..5695)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5886..5905
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6190..6212
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6204..6221
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6205..6221
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6241..6598)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6255..6390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6407..6598
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6555..6575
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    6667..6801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6691..6710)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6745..6764
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      6805..6814
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gctgaaagac cggccaaagg ggcccttggg gtcctcgcct ggccgcaggc cagggcccaa
       61 cggctgacca tggaagggtc ccacctccga gaggtcgggc agcgtccggt ttcgaggggg
      121 cgtcatcacc acgccctgcc gggctaggag ggccagcagg ttctgggagc tgggggtctt
      181 cgggaagtca aaggagggca cagccttggt gggggagccc aggatggggg gcagggggtt
      241 tcgctgcagg aagtcgggca gcttgggtga gccccgcagg ttccggcagg actgcaggcc
      301 gtgggacggc tggggagggc tggcctgtgg tgggctgaac acagctccca gggggtccgt
      361 ggggggtttg ggcagcttgg ggcggacgac gtgcaagtca gacaggttgg gggcgctgtg
      421 caggcggcag cccagcccgg aagtgcgggg agagtgctcg ggtgcagagg agcctggacg
      481 aggggacctg ccaccccgca tctcagctcc ctgtggggag ggcgtcccgc tccagcctgg
      541 ccgctcaggg atggttccaa cttgaggaga tggcgtgtag ggccggcctc cacccagaga
      601 catcttcctg gccaggacgc ctccatgctc agcgtgggca gggggcgagg ggctggccct
      661 tgcaaagccc agggggctgg tgctgcctga cctgcggatg gcagaggacc gaggtgtttg
      721 gaactgggtg ggtgactgca ggtttcgctc aatgcgctgg tagttctgca cctgcgtggg
      781 gactgggatg gggacagagg cgccgcacct gctgctggag aacgggccag cccggcctga
      841 gggactgggg gagctgctgc agggtgggga tggagatggg gtccggccgt ggctctccaa
      901 gcccgcagag gccaccagtg agctcccact gcacatcagg ctgtctggcg ggggtttggc
      961 actgggcgcc tcagccacca ggtcacctgg aaactgcgcg gggaccatga cgaagtcgtc
     1021 tgtgtcacag gaagagtcct tgctgccacc agagtcccgg gagctgtgca ggaagccagc
     1081 ggtgtcagcc ggggaggcca gggtcttctg cagctgctgc atctcgccca gggacggcgg
     1141 ggaggccagg tgggaggtgg agctgctgct ggagctgctg ccggaccccg agcttgggta
     1201 cgagggcaca ggcacgggtg gggatttcct gaccgagggg ctggcatcga ggaaagggtg
     1261 atgaaaaaac tcatcgaagt ccatgcggtc cttgtggttg cgttgcagta gggccaggag
     1321 cagctgccgc agcggggccg aggtctcccg ggggatggtg gggaccaacg tcttgttctt
     1381 ctcgtagaac aggcgcaggt cctgggggct gctggcctgg aagggcgcct tccccgtcag
     1441 gcactggtag acgatggtgc cgatgctcca caggtccgcc ttcccgtcgt agtgctggga
     1501 catgatgacc tcgggggcca tgtacatggg ggagccgcag agtgtggccg ccatcatgtt
     1561 gctctggagg taccgcgcga agccgaagtc agcgatcttg acgcggatgc tgttggggtt
     1621 ggcgcggcgg ccggcggggt tggacagcag gatgttctgc ggtttcaggt cgcggtggat
     1681 gatgcctttg ctgtgcagaa gccgcatggc gcccgcgatc tgctgcagga agagcctgat
     1741 ggtgtcctcg ctcagcgtgc gcatggcgtg caggtagtcg gccaggtccc caccgttgca
     1801 gtactccata accaggtaga cagaattagc catttcctgg aagtcgtaca gggccacgat
     1861 gttttcatgt ttcagttcct tcaggatttt gatttccttc cccagcagcg tctgagactt
     1921 ggcgaggttc ttcttgttaa tgcacttgac ggcgacctcc aaatcgtgct tctcgcggtg
     1981 gcggcccttg aagaccaccg cgaaggcgcc gtggccgatc aggtccttgc gggagaactc
     2041 gaacttgccc acggtctctg tgccgccgcg gccgggctcc atggtcgacc caccaccacc
     2101 accattgaga tctgcgtagt caggcacatc ataggggtag cccatggtgg cgcgaatggc
     2161 cttggtgaag agaccgactg tcggttcggg aattcaatcg atagaaccga ggtgcagttg
     2221 gacctgggag tggacacctg tggagagaaa ggcaaagtgg atgttattct atagtgtcac
     2281 ctaaatcgta tgtgtatgat acataaggtt atgtattaat tgtagccgcg ttctaacgaa
     2341 gccaaggggg tgggcctata gactctatag gcggtactta cgtcactctt ggcacgggga
     2401 atccgcgttc caatgcaccg ttcccggccg cggaggctgg atcggtcccg gtgtcttcta
     2461 tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac taaacgagct
     2521 ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag
     2581 ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg
     2641 tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg
     2701 tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa
     2761 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     2821 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     2881 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     2941 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     3001 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     3061 gtaattgatt actattaata actagtcaat aatcaatgtc gggcgagctc gaattaattc
     3121 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3181 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3241 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3301 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3361 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3421 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3481 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3541 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3601 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3661 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3721 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3781 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3841 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3901 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3961 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4021 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4081 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4141 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4201 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4261 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4321 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4381 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4441 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4501 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4561 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4621 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4681 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4741 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4801 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4861 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4921 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4981 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5041 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5101 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5161 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5221 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5281 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5341 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5401 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5461 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5521 ctgagagtgc accataaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     5581 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5641 aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5701 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     5761 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     5821 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     5881 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5941 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtactatggt
     6001 tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6061 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     6121 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     6181 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg ttaacagctt
     6241 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     6301 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     6361 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     6421 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     6481 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     6541 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagaa
     6601 ggtacctaac caagttcctc tttcagaggt tatttcaggc catggtgctg cgccgaatta
     6661 attcccgatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     6721 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     6781 ataagctgca ataaacaagt tgggccatgg cggccaagct ggggatcctt acttacttag
     6841 cggccgctca ggcacagatg ccagtcagca gcgccgagag tctccgctca atgcacagct
     6901 tgcacttggt gacgttctcg atgtcggcct ggtccgagag catgtgctgc agcccctcca
     6961 ggagcagcag ggccttgtgg tagcgtggga cgcagccctc acggtgctgg aacatctcgt
     7021 ccagggcagc cgactgcacc atctgcacag cgtggctgaa gatgagcctc tcggcagtga
     7081 tgctgtgaat gcggtccagg agccgctgct tgtccaggaa gaagcgctgc agccgcaggc
     7141 tcaggccctg gcaggacacc acgctggcct tgtacagctc attcagcctg cgcaccacct
     7201 gcttcacagt ggacgacagg cagagcttgc cggcccggat ctggtcgatg gcactttgca
     7261 ggccggagga cagtagctcg gccaccttca ggtacagcac cagctgttcc gcgaagcccc
     7321 attctcggct cagcaggctg atctggtcgg ccaccacact ctcctgcagc tggtactcag
     7381 ggccccccgc cgcctcactg gcgctgccct tcagggctgc gatctccagg acgtgctgca
     7441 cgaacagcag cgtgaagcgc aggccacgca ggatctccgt gtgctcttgc tccatgaggg
     7501 tctcctcagg gaggtcgggg gcctcgaagg tcacagcccc ctccaggttc gcagcaatgg
     7561 ggtcggcaaa gctgcagccg tgtcctggag cagggagctc aggggctggg gcctcgctgc
     7621 agggcccagg caccaggtgg cgggcagaag agctggcaga gccagtgggg cccgctgaga
     7681 acatcctggt gcgggggccc tgggggggcg tgctcccgct cgggggagag cccacggtga
     7741 agaccaccgg ggaggggctg ctggtgcccc cagcacgggc tcctgggtgc aggctccctc
     7801 caaagccagc tgagggtgcg atctccatgg gcttctcctg caggctctcc gtgctgcccg
     7861 ggtccggggc ttgtgtccca aacgccgcct taaggagcag gtcagtgagg cggctggt
//
