LOCUS       sequence-451466-        5586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express human SKP1 in insect cells, such as Trichoplusia ni Hi5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5586)
  AUTHORS   Cao S, Garcia SF, Shi H, James EI, Kito Y, Shi H, Mao H, Kaisari S, 
            Rona G, Deng S, Goldberg HV, Ponce J, Ueberheide B, Lignitto L, 
            Guttman M, Pagano M, Zheng N
  TITLE     Recognition of BACH1 quaternary structure degrons by two F-box 
            proteins under oxidative stress.
  JOURNAL   Cell. 2024 Oct 30:S0092-8674(24)01188-7. doi: 
            10.1016/j.cell.2024.10.012.
  PUBMED    39504958
REFERENCE   2  (bases 1 to 5586)
  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: "Cell. 2024 
            Oct 30:S0092-8674(24)01188-7. doi: 10.1016/j.cell.2024.10.012."
COMMENT     Sequence Label: pFB-His-SKP1
FEATURES             Location/Qualifiers
     source          1..5586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  140..305
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      489..943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(576..595)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     785..806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        969..1073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1074..1142,1143..1934)
                     /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(1292..1311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2105..2693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2594..2613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2847..2864
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2998..3222
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3289..3822)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3992..3997,3998..4014,4015..4020))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        4368..4459
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4384..4401
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4526..4543
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4565..4585
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    5250..5384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5287..5306)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5341..5360
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 acgagcaaga tcctcgccaa ggtgatcgag tattgcaaga ggcacgtcga ggctgctgcc
       61 tctaaggccg aggccgtcga gggtgctgct acctccgatg acgatcttaa ggcctggcct
      121 gggcggccta ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt
      181 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
      241 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
      301 ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg
      361 tgacaaaccg tcatcttcgg ctactttttc tctgtcacag aatgaaaatt tttctgtcat
      421 ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttgca gcctgaatgg
      481 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      541 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      601 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      661 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      721 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      781 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
      841 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
      901 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
      961 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggc tctcgcggta
     1801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2221 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata gaccagccgc
     2941 gtaacctggc aaaatcggtt acggttgagt aataaatgga tgccctgcgt aagcgggtgt
     3001 gggcggacaa taaagtctta aactgaacaa aatagatcta aactatgaca ataaagtctt
     3061 aaactagaca gaatagttgt aaactgaaat cagtccagtt atgctgtgaa aaagcatact
     3121 ggacttttgt tatggctaaa gcaaactctt cattttctga agtgcaaatt gcccgtcgta
     3181 ttaaagaggg gcgtggccaa gggcatggta aagactatat tcgcggcgtt gtgacaattt
     3241 accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt aggtggcggt
     3301 acttgggtcg atatcaaagt gcatcacttc ttcccgtatg cccaactttg tatagagagc
     3361 cactgcggga tcgtcaccgt aatctgcttg cacgtagatc acataagcac caagcgcgtt
     3421 ggcctcatgc ttgaggagat tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc
     3481 ggagactgcg agatcataga tatagatctc actacgcggc tgctcaaact tgggcagaac
     3541 gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt
     3601 cttactacgg agcaagttcc cgaggtaatc ggagtccggc tgatgttggg agtaggtggc
     3661 tacgtctccg aactcacgac cgaaaagatc aagagcagcc cgcatggatt tgacttggtc
     3721 agggccgagc ctacatgtgc gaatgatgcc catacttgag ccacctaact ttgttttagg
     3781 gcgactgccc tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc
     3841 gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca
     3901 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     3961 gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa
     4021 ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag
     4081 cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgc tctccacgca
     4141 tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc
     4201 ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc
     4261 ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga
     4321 ctctagctat agttctagtg gttggctacg taccggaata ttaatagatc atggagataa
     4381 ttaaaatgat aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt
     4441 aataaaaaaa cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc
     4501 tcggtccgaa accatgtcgt actaccatca ccatcaccat cacgattacg atatcccaac
     4561 gaccgaaaac ctctacttcc aatcgatgcc ttcaattaag ttgcagagtt ctgatggaga
     4621 gatatttgaa gttgatgtgg aaattgccaa acaatctgtg actattaaga ccatgttgga
     4681 agatttggga atggatgatg aaggagatga tgacccagtt cctctaccaa atgtgaatgc
     4741 agcaatatta aaaaaggtca ttcagtggtg cacccaccac aaggatgacc ctcctcctcc
     4801 tgaagatgat gagaacaaag aaaagcgaac agatgatatc cctgtttggg accaagaatt
     4861 cctgaaagtt gaccaaggaa cactttttga actcattctg gctgcaaact acttagacat
     4921 caaaggtttg cttgatgtta catgcaagac tgttgccaat atgatcaagg ggaaaactcc
     4981 tgaggagatt cgcaagacct tcaatatcaa aaatgacttt actgaagagg aggaagccca
     5041 ggtacgcaaa gagaaccagt ggtgtgaaga gaagtgaccg gaggatgagt gtggcctgca
     5101 gtctcgaggc atgcggtacc aagcttgtcg agaagtacta gaggatcata atcagccata
     5161 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     5221 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     5281 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5341 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagt
     5401 aatcccacca ctcctctcca cttaattaac ctagaggccg cgctggcctc tgcgaagaag
     5461 attgtgttga agagttccga tggtgaatct ttcgaggttg aggaggcggt ggctctcgag
     5521 tcacaaacca tagcgcatat ggttgaagac gactgcgtcg acaacggagt ccctcttcct
     5581 aacgtc
//
