LOCUS       sequence-490909-        7987 bp    DNA     circular SYN 28-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7987)
  AUTHORS   Kleijnen MF, Shih AH, Zhou P, Kumar S, Soccio RE, Kedersha NL, Gill 
            G, Howley PM
  TITLE     The hPLIC proteins may provide a link between the ubiquitination 
            machinery and the proteasome.
  JOURNAL   Mol Cell 2000 Aug;6(2):409-19.
  PUBMED    10983987
REFERENCE   2  (bases 1 to 7987)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell"; 
            date: "2000-08"; volume: "6(2)"; pages: "409-19"
COMMENT     Sequence Label: p4455 FLAG-hPLIC-2
FEATURES             Location/Qualifiers
     source          1..7987
                     /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"
     CDS             682..705
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3838..4460
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4037..4056)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4484..4504)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4489..4818
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4669..4804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4857..4953
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     primer_bind     complement(5029..5048)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5030..5048)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5062..5078)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5062..5078)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5075..5097)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5086..5102
                     /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(5110..5116,5117..5134,5135..5140))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5155..5176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5293..5310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5464..6052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5544..5563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6223..7014,7015..7083))
                     /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     6846..6865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7084..7188)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7215..7670
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7302..7321)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7512..7533
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7796..7818
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7810..7827
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7811..7827
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgttt ttatttttaa ttttctttca aatacttcca tcgaattcag atctggtacc
      661 acgcgtatcg ataagcttat ggactacaag gacgacgatg acaaggctga gaatggcgag
      721 agcagcggcc ccccgcgccc ctcccgcggc cctgctgcgg cccaaggctc ggctgctgcc
      781 ccggctgagc ctaaaatcat caaagtcacg gtgaagactc ccaaagagaa agaggagttc
      841 gcggtgcccg agaacagctc ggttcagcag tttaaggaag cgatttcgaa acgcttcaaa
      901 tcccaaaccg atcagctagt gctgattttt gccggaaaaa tcttaaaaga tcaagatacc
      961 ttgatccagc atggcatcca tgatgggctg actgttcacc ttgtcatcaa aagccagaac
     1021 cgacctcagg gccagtccac gcagcctagc aatgccgcgg gaactaacac tacctcggcg
     1081 tcgactccca ggagtaactc cacacctatt tccacaaata gcaacccgtt tgggttgggg
     1141 agcctgggag gacttgcagg ccttagcagc ctgggcttga gctcgaccaa cttctctgag
     1201 ctccagagcc agatgcagca gcagcttatg gccagccctg agatgatgat ccaaataatg
     1261 gaaaatccct ttgttcagag catgctttcg aatcccgatc tgatgaggca gctcattatg
     1321 gctaatccac agatgcagca attgattcag agaaacccag aaatcagtca cctgctcaac
     1381 aacccagaca taatgaggca gacactcgaa attgccagga atccagccat gatgcaagag
     1441 atgatgagaa atcaagacct ggctcttagc aatctagaaa gcatcccagg tggctataat
     1501 gctttacggc gcatgtacac tgacattcaa gagccgatgc tgaatgccgc acaagagcag
     1561 tttgggggta atccatttgc ctccgtgggg agtagttcct cctctgggga aggtacgcag
     1621 ccttcccgca cagaaaatcg cgatccacta cccaatccat gggcaccacc gccagctacc
     1681 cagagttctg caactaccag cacgaccaca agcactggta gtgggtctgg caatagttcc
     1741 agcaatgcta ctgggaacac cgttgctgcc gctaattatg tcgccagcat ctttagtacc
     1801 ccaggcatgc agagcctgct gcaacagata actgaaaacc cccagctgat tcagaatatg
     1861 ctgtcggcgc cctacatgag aagcatgatg cagtcgctga gccagaatcc agatttggct
     1921 gcacagatga tgctgaatag cccgctgttt actgcaaatc ctcagctgca ggagcagatg
     1981 cggccacagc tcccagcctt cctgcagcag atgcagaatc cagacacact atcagccatg
     2041 tcaaacccaa gagcaatgca ggctttaatg cagatccagc aggggctaca gacattagcc
     2101 actgaagcac ctggcctgat tccgagcttc actccaggtg tgggggtggg ggtgctggga
     2161 accgctatag gccctgtagg cccagtcacc cccataggcc ccataggccc tatagtccct
     2221 tttaccccca taggccccat tgggcccata ggacccactg gccctgcagc cccccctggc
     2281 tccaccggct ctggtggccc cacggggcct actgtgtcca gcgctgcacc tagtgaaacc
     2341 acgagtccta catcagaatc tggacccaac cagcagttca ttcagcaaat ggtgcaggcc
     2401 ctggctggag caaatgctcc acagctgccg aatccagaag tcagatttca gcaacaactg
     2461 gaacagctca acgcaatggg gttcttaaac cgtgaagcaa acttgcaggc cctaatagca
     2521 acaggaggcg acatcaatgc agccattgaa aggctgctgg gctcccagcc atcgtaatca
     2581 catttctgta cctggaaaaa aaatgtatct tatttttgat aatggctctt aaatctttaa
     2641 acacacacac aaaatcgttc tttactttca ttttgattct tttaaatctg tctagttgta
     2701 agtctaatat gatgcatttt aagatggagt ccctccctcc tacttccctc actccctttc
     2761 tcctttgctt atttttccta ccttcccttc ctcttgtctc cccactccct ccctctttgt
     2821 ttccttcctt ccttatttcc tttagtttcc ttccttagcc gttttgagtg gtgggaatca
     2881 atgctgtttc actcaaaagt gttgcatgca aacacttctc tttattctgc atttattgtg
     2941 atttttggaa acaggtatca accttcacag ttgggtgaac aagtgttgtc ctacagatgt
     3001 ccaatttatt tgcattttta aacattagcc tatgatagta atttaatgta gaatgaagat
     3061 attaaaaaca gaagcaaatt atttgaagct ctctaatttg tggtacgata ttgcttattg
     3121 tgactttggc atgtattttt gctagcaaaa tgctgtaaga tttataccat tgatcttttt
     3181 tgctatattt gtatacagta cagtaagcac aattggcact gtacatctaa aaatattaca
     3241 gtagaatctg agtgtaatat gtgtaaccaa aatgagaaag aatacaagaa atgtttctgg
     3301 agctagttat gtctcacaat tttgtagaat cttacagcat ctttgataaa cttctcagtg
     3361 aaaatgttgg ctaggcaagt tcagttaaaa tatagtagaa atgtttatcc tggtatctct
     3421 aagtatacat ttaattgtac agaaaattta cagtgtaaca ttgtgtcaac atttgcagat
     3481 tgactgtata tgaccttaat ctttgtgcag cctgaaggat cagtgtagta atgccaggaa
     3541 agtgcttttt acctaagact tccttctcag cttctcccat aaagagaccc taatatgcat
     3601 tttgatttgt aattggaaat gtaactttca ctgaaagtgt catgtgatgt ttgcattact
     3661 tttaactgct atgtataaag gaaagtgtgt cttttgactt catcagttat ttctcttgtg
     3721 cacagagaaa aatgcattaa aaatgactaa aaaaaataaa aaattaaaaa atggaaaaaa
     3781 aaaaaaaaaa aaaaaaaaac tcgagactag tgcggccgct cgagtctaga ggatcccggg
     3841 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3901 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3961 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     4021 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     4081 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     4141 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     4201 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     4261 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     4321 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     4381 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     4441 cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta
     4501 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4561 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4621 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4681 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4741 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4801 ggcctaggct tttgcaaaaa gctcctcgag gaactgaaaa accagaaagt taactggtaa
     4861 gtttagtctt tttgtctttt atttcaggtc ccggatccgg tggtggtgca aatcaaagaa
     4921 ctgctcctca gtggatgttg cctttacttc taggcctgta cggaagtgtt acttctgctc
     4981 taaaagctgc tgcaggtcga ctctagggcc gcgtaccgag ctcgaattcc ctatagtgag
     5041 tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5101 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5161 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5221 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5281 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5341 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5401 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5881 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6181 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6241 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6301 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6361 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6421 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6481 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6541 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6601 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6661 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6721 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6781 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6841 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6901 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6961 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7021 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7081 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7141 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7201 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     7261 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     7321 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     7381 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     7441 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     7501 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     7561 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     7621 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca
     7681 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     7741 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     7801 tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg
     7861 caattagcca tattagtcat tggttatata gcataaatca atattggcta ttggccattg
     7921 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
     7981 ccatgtt
//
