LOCUS       sequence-42-expo        7236 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 7236)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7236)
  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"
COMMENT     Sequence Label: pOK11_34
FEATURES             Location/Qualifiers
     source          1..7236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             324..344
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             432..3479
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(434..456)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4810..4832)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4932..4951
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5029..5047)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5115..5219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5220..5288,5289..6080)
                     /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(5438..5457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6251..6839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6740..6759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6993..7010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7127..7148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7163..7168,7169..7186,7187..7193)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7201..7217
                     /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     7206..7228
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7225..7236,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7225..7236,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggcctt ggtcgactct agacactttt
       61 cagctaccta gatacatgga tatccccgcc tcccaatcca cccacccagg gaaaaagaag
      121 ggctcgccga aaatcaaagt tatctccagg ctcgcgcatc ccaccgagcg gttgacttct
      181 ctccaccact tttcatttta accctcgggg tacgggattg gccaaaggac ccaaaggtat
      241 gtttcgaatg atactaacat aacatagaac attttcagga ggacccttct tggagggtac
      301 cgagctcaga aaaaatgact gctccaaaga agaagcgtaa ggtaccggtg ggtgaagacc
      361 agaaacagca cctcgaactg agccgcgata ttgcccagcg tttcaacgcg ctgtatggcg
      421 agatcgatcc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      481 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      541 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg
      601 caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg
      661 tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtgacct
      721 atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc
      781 tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg
      841 gcgttaactc ggcgtttcat ctgtggtgca acgggcgctg ggtcggttac ggccaggaca
      901 gtcgtttgcc gtctgaattt gacctgagcg catttttacg cgccggagaa aaccgcctcg
      961 cggtgatggt gctgcgctgg agtgacggca gttatctgga agatcaggat atgtggcgga
     1021 tgagcggcat tttccgtgac gtctcgttgc tgcataaacc gactacacaa atcagcgatt
     1081 tccatgttgc cactcgcttt aatgatgatt tcagccgcgc tgtactggag gctgaagttc
     1141 agatgtgcgg cgagttgcgt gactacctac gggtaacagt ttctttatgg cagggtgaaa
     1201 cgcaggtcgc cagcggcacc gcgcctttcg gcggtgaaat tatcgatgag cgtggtggtt
     1261 atgccgatcg cgtcacacta cgtctgaacg tcgaaaaccc gaaactgtgg agcgccgaaa
     1321 tcccgaatct ctatcgtgcg gtggttgaac tgcacaccgc cgacggcacg ctgattgaag
     1381 cagaagcctg cgatgtcggt ttccgcgagg tgcggattga aaatggtctg ctgctgctga
     1441 acggcaagcc gttgctgatt cgaggcgtta accgtcacga gcatcatcct ctgcatggtc
     1501 aggtcatgga tgagcagacg atggtgcagg atatcctgct gatgaagcag aacaacttta
     1561 acgccgtgcg ctgttcgcat tatccgaacc atccgctgtg gtacacgctg tgcgaccgct
     1621 acggcctgta tgtggtggat gaagccaata ttgaaaccca cggcatggtg ccaatgaatc
     1681 gtctgaccga tgatccgcgc tggctaccgg cgatgagcga acgcgtaacg cgaatggtgc
     1741 agcgcgatcg taatcacccg agtgtgatca tctggtcgct ggggaatgaa tcaggccacg
     1801 gcgctaatca cgacgcgctg tatcgctgga tcaaatctgt cgatccttcc cgcccggtgc
     1861 agtatgaagg cggcggagcc gacaccacgg ccaccgatat tatttgcccg atgtacgcgc
     1921 gcgtggatga agaccagccc ttcccggctg tgccgaaatg gtccatcaaa aaatggcttt
     1981 cgctacctgg agagacgcgc ccgctgatcc tttgcgaata cgcccacgcg atgggtaaca
     2041 gtcttggcgg tttcgctaaa tactggcagg cgtttcgtca gtatccccgt ttacagggcg
     2101 gcttcgtctg ggactgggtg gatcagtcgc tgattaaata tgatgaaaac ggcaacccgt
     2161 ggtcggctta cggcggtgat tttggcgata cgccgaacga tcgccagttc tgtatgaacg
     2221 gtctggtctt tgccgaccgc acgccgcatc cagcgctgac ggaagcaaaa caccagcagc
     2281 agtttttcca gttccgttta tccgggcaaa ccatcgaagt gaccagcgaa tacctgttcc
     2341 gtcatagcga taacgagctc ctgcactgga tggtggcgct ggatggtaag ccgctggcaa
     2401 gcggtgaagt gcctctggat gtcgctccac aaggtaaaca gttgattgaa ctgcctgaac
     2461 taccgcagcc ggagagcgcc gggcaactct ggctcacagt acgcgtagtg caaccgaacg
     2521 cgaccgcatg gtcagaagcc gggcacatca gcgcctggca gcagtggcgt ctggcggaaa
     2581 acctcagtgt gacgctcccc gccgcgtccc acgccatccc gcatctgacc accagcgaaa
     2641 tggatttttg catcgagctg ggtaataagc gttggcaatt taaccgccag tcaggctttc
     2701 tttcacagat gtggattggc gataaaaaac aactgctgac gccgctgcgc gatcagttca
     2761 cccgtgcacc gctggataac gacattggcg taagtgaagc gacccgcatt gaccctaacg
     2821 cctgggtcga acgctggaag gcggcgggcc attaccaggc cgaagcagcg ttgttgcagt
     2881 gcacggcaga tacacttgct gatgcggtgc tgattacgac cgctcacgcg tggcagcatc
     2941 aggggaaaac cttatttatc agccggaaaa cctaccggat tgatggtagt ggtcaaatgg
     3001 cgattaccgt tgatgttgaa gtggcgagcg atacaccgca tccggcgcgg attggcctga
     3061 actgccagct ggcgcaggta gcagagcggg taaactggct cggattaggg ccgcaagaaa
     3121 actatcccga ccgccttact gccgcctgtt ttgaccgctg ggatctgcca ttgtcagaca
     3181 tgtatacccc gtacgtcttc ccgagcgaaa acggtctgcg ctgcgggacg cgcgaattga
     3241 attatggccc acaccagtgg cgcggcgact tccagttcaa catcagccgc tacagtcaac
     3301 agcaactgat ggaaaccagc catcgccatc tgctgcacgc ggaagaaggc acatggctga
     3361 atatcgacgg tttccatatg gggattggtg gcgacgactc ctggagcccg tcagtatcgg
     3421 cggaattcca actgagcgcc ggtcgctacc attaccaact tgtctggtgt caaaaataat
     3481 aggggccgct gtcatcagat cgccatctcg cgcccgtgcc tctgacttct aagtccaatt
     3541 actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt
     3601 atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat
     3661 gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc
     3721 tccctccccc cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt
     3781 cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa
     3841 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct
     3901 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat
     3961 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc
     4021 cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     4081 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     4141 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     4201 tacattccca atataccaaa cataactgtt taaaattaaa catttttcta aattttatat
     4261 gatttctttt aaatttgcaa aaattactta aatttgaatt cccgcgcaaa tgagtgactt
     4321 cattttctgc attattgtgt tttccggcta tattaatagg tatttgtttg tgtttttctt
     4381 tattttatga ttcgaactcc aatttgtaaa ttttcgaaca tatttcccta aagaaaaaat
     4441 atgattaatc tggaaaaatt ggaaaattat ttttcaaata aaaaacaaag aaaaaaatga
     4501 agaaaaacct attagtttgg ccataaaacg caaaaatgtc gaaaatgacg tcactcatct
     4561 gcgcgggaaa tcaagaataa ttcggccttt tttatttttt tggaaaatcg taaaacattt
     4621 agaaaaattt tttaatagtt atatgggact tattctgcat ttagggcaaa agccagagac
     4681 tactccaccg ttaacatgaa ttatgaatat tattgcgaca agacccaaac attgataaac
     4741 ccaaatctag cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga
     4801 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4861 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4921 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     4981 accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc ctcgtgatac
     5041 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5101 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5161 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5221 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5281 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5341 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5401 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5461 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5521 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5581 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5641 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5701 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5761 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5821 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5881 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5941 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6001 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6061 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6121 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6181 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6361 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6421 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6481 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6541 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6601 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     6661 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6721 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6781 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6841 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6901 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6961 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7021 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     7081 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7141 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7201 ttgtgagcgg ataacaattt cacacaggaa acagct
//
