LOCUS       sequence-157862-        6330 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 6330)
  AUTHORS   Michel JJ, Xiong Y
  TITLE     Human CUL-1, but not other cullin family members, selectively 
            interacts with SKP1 to form a complex with SKP2 and cyclin A.
  JOURNAL   Cell Growth Differ. 1998 Jun . 9(6):435-49.
  PUBMED    9663463
REFERENCE   2  (bases 1 to 6330)
  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 Growth
            Differ. 1998 Jun . 9(6):435-49."
COMMENT     Sequence Label: pGEX-Skp2
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        63..140
                     /gene="lacI"
                     /label=lacI promoter
     CDS             141..1223
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(160..179)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1272..1277,1278..1295,1296..1302)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1310..1326
                     /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)."
     primer_bind     1334..1350
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1334..1350
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1346..1873
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1366..1383)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1366..1382)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1375..1397)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1975..1980,1981..1996,1997..2003)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2011..2027
                     /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)."
     CDS             2050..2703
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2661..2683
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2710..2727
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4172..4194)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4339..4357)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4425..4529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4530..4598,4599..5390)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4748..4767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5561..6149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6050..6069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6303..6320
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
       61 ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
      121 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      181 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      241 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      301 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      361 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      421 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      481 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      541 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      601 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      661 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      721 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      781 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      841 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      901 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
      961 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     1021 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1081 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1141 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1201 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1261 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     1321 taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt
     1381 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     1441 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     1501 gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga
     1561 aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca
     1621 gatgcacggt tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc
     1681 gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga
     1741 aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc
     1801 gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct
     1861 gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
     1921 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca
     1981 attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     2041 acagtattca tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact
     2101 cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa
     2161 ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat
     2221 tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac
     2281 aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga
     2341 gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact
     2401 ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta
     2461 tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac
     2521 gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt
     2581 tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa atccagcaag
     2641 tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca
     2701 aaatcggatc tggttccgcg tggatccccg ggaatttccg gtggtggtgg tggaattcta
     2761 gactccatgg acgtatttaa aactcccggg cctgcggacg ctatgcacag gaagcacctc
     2821 caggagattc cagacctgag tagcaacgtt gccaccagct tcacgtgggg atgggattcc
     2881 agcaagactt ctgaactgct gtcaggcatg ggggtctccg ccctggagaa agaggagccc
     2941 gacagtgaga acatccccca ggaactgctc tcaaacctgg gccacccgga gagcccccca
     3001 cggaaacggc tgaagagcaa agggagtgac aaagactttg tgattgtccg caggcctaag
     3061 ctaaatcgag agaactttcc aggtgtttca tgggactccc ttccggatga gctgctcttg
     3121 ggaatctttt cctgtctgtg cctccctgag ctgctaaagg tctctggtgt ttgtaagagg
     3181 tggtatcgcc tagcgtctga tgagtctcta tggcagacct tagacctcac aggtaaaaat
     3241 ctgcacccgg atgtgactgg tcggttgctg tctcaagggg tgattgcctt ccgctgccca
     3301 cgatcattta tggaccaacc attggctgaa catttcagcc cttttcgtgt acagcacatg
     3361 gacctatcga actcagttat agaagtgtcc accctccacg gcatactgtc tcagtgttcc
     3421 aagttgcaga atctaagcct ggaaggcctg cggctttcgg atcccattgt caatactctc
     3481 gcaaaaaact caaatttagt gcgacttaac ctttctgggt gttctggatt ctctgaattt
     3541 gccctgcaga ctttgctaag cagctgttcc agactggatg agctgaacct ctcctggtgt
     3601 tttgatttca ctgaaaagca tgtacaggtg gctgttgcgc atgtgtcaga gaccatcacc
     3661 cagctgaatc ttagcggcta cagaaagaat ctccagaaat cagatctctc tactttagtt
     3721 agaagatgcc ccaatcttgt ccatctagac ttaagtgata gtgtcatgct aaagaatgac
     3781 tgctttcagg aaattttcca gctcaactac ctccaacacc tatcactcag tcggtgctat
     3841 gatataatac ctgaaacttt acttgaactt ggagaaattc ccacactaaa aacactacaa
     3901 gtttttggaa tcgtgccaga tggtaccctt caactgttaa aggaagccct tcctcatcta
     3961 cagattaatt gctcccattt caccaccatt gccaggccaa ctattggcaa caaaaagaac
     4021 caggagatat ggggcatcaa atgccgactg acactgcaaa agcccagttg tctatgaagt
     4081 atttattgca ggatggtgtc tcgagctcaa gcttaattca tcgtgactga ctgacgatct
     4141 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4201 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4261 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     4321 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4381 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4441 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4501 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4561 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4621 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4681 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4741 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     4801 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4861 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4921 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4981 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5041 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     5101 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5161 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5221 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5281 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5341 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5401 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5461 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5521 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5581 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5641 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5701 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5761 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5821 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5881 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5941 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6001 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6061 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6121 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6181 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6241 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6301 gcagcgagtc agtgagcgag gaagcggaag
//
