LOCUS       sequence-52-expo        7242 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 7242)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7242)
  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: pPD26_77
FEATURES             Location/Qualifiers
     source          1..7242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             85..105
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             193..3240
                     /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(195..217)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     RBS             3464..3472
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(4529..4547)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4568..4584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4592..4608
                     /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(4616..4622,4623..4640,4641..4646))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4816..4838)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4938..4957
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5035..5053)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5121..5225
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5226..5294,5295..6086)
                     /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(5444..5463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6257..6845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6746..6765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6999..7016
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7133..7154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7169..7174,7175..7192,7193..7199)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7207..7223
                     /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     join(7231..7242,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta
       61 ccgagctcag aaaaaatgac tgctccaaag aagaagcgta aggtaccggt gggtgaagac
      121 cagaaacagc acctcgaact gagccgcgat attgcccagc gtttcaacgc gctgtatggc
      181 gagatcgatc ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
      241 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
      301 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg
      361 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc
      421 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc
      481 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg
      541 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat
      601 ggcgttaact cggcgtttca tctgtggtgc aacgggcgct gggtcggtta cggccaggac
      661 agtcgtttgc cgtctgaatt tgacctgagc gcatttttac gcgccggaga aaaccgcctc
      721 gcggtgatgg tgctgcgctg gagtgacggc agttatctgg aagatcagga tatgtggcgg
      781 atgagcggca ttttccgtga cgtctcgttg ctgcataaac cgactacaca aatcagcgat
      841 ttccatgttg ccactcgctt taatgatgat ttcagccgcg ctgtactgga ggctgaagtt
      901 cagatgtgcg gcgagttgcg tgactaccta cgggtaacag tttctttatg gcagggtgaa
      961 acgcaggtcg ccagcggcac cgcgcctttc ggcggtgaaa ttatcgatga gcgtggtggt
     1021 tatgccgatc gcgtcacact acgtctgaac gtcgaaaacc cgaaactgtg gagcgccgaa
     1081 atcccgaatc tctatcgtgc ggtggttgaa ctgcacaccg ccgacggcac gctgattgaa
     1141 gcagaagcct gcgatgtcgg tttccgcgag gtgcggattg aaaatggtct gctgctgctg
     1201 aacggcaagc cgttgctgat tcgaggcgtt aaccgtcacg agcatcatcc tctgcatggt
     1261 caggtcatgg atgagcagac gatggtgcag gatatcctgc tgatgaagca gaacaacttt
     1321 aacgccgtgc gctgttcgca ttatccgaac catccgctgt ggtacacgct gtgcgaccgc
     1381 tacggcctgt atgtggtgga tgaagccaat attgaaaccc acggcatggt gccaatgaat
     1441 cgtctgaccg atgatccgcg ctggctaccg gcgatgagcg aacgcgtaac gcgaatggtg
     1501 cagcgcgatc gtaatcaccc gagtgtgatc atctggtcgc tggggaatga atcaggccac
     1561 ggcgctaatc acgacgcgct gtatcgctgg atcaaatctg tcgatccttc ccgcccggtg
     1621 cagtatgaag gcggcggagc cgacaccacg gccaccgata ttatttgccc gatgtacgcg
     1681 cgcgtggatg aagaccagcc cttcccggct gtgccgaaat ggtccatcaa aaaatggctt
     1741 tcgctacctg gagagacgcg cccgctgatc ctttgcgaat acgcccacgc gatgggtaac
     1801 agtcttggcg gtttcgctaa atactggcag gcgtttcgtc agtatccccg tttacagggc
     1861 ggcttcgtct gggactgggt ggatcagtcg ctgattaaat atgatgaaaa cggcaacccg
     1921 tggtcggctt acggcggtga ttttggcgat acgccgaacg atcgccagtt ctgtatgaac
     1981 ggtctggtct ttgccgaccg cacgccgcat ccagcgctga cggaagcaaa acaccagcag
     2041 cagtttttcc agttccgttt atccgggcaa accatcgaag tgaccagcga atacctgttc
     2101 cgtcatagcg ataacgagct cctgcactgg atggtggcgc tggatggtaa gccgctggca
     2161 agcggtgaag tgcctctgga tgtcgctcca caaggtaaac agttgattga actgcctgaa
     2221 ctaccgcagc cggagagcgc cgggcaactc tggctcacag tacgcgtagt gcaaccgaac
     2281 gcgaccgcat ggtcagaagc cgggcacatc agcgcctggc agcagtggcg tctggcggaa
     2341 aacctcagtg tgacgctccc cgccgcgtcc cacgccatcc cgcatctgac caccagcgaa
     2401 atggattttt gcatcgagct gggtaataag cgttggcaat ttaaccgcca gtcaggcttt
     2461 ctttcacaga tgtggattgg cgataaaaaa caactgctga cgccgctgcg cgatcagttc
     2521 acccgtgcac cgctggataa cgacattggc gtaagtgaag cgacccgcat tgaccctaac
     2581 gcctgggtcg aacgctggaa ggcggcgggc cattaccagg ccgaagcagc gttgttgcag
     2641 tgcacggcag atacacttgc tgatgcggtg ctgattacga ccgctcacgc gtggcagcat
     2701 caggggaaaa ccttatttat cagccggaaa acctaccgga ttgatggtag tggtcaaatg
     2761 gcgattaccg ttgatgttga agtggcgagc gatacaccgc atccggcgcg gattggcctg
     2821 aactgccagc tggcgcaggt agcagagcgg gtaaactggc tcggattagg gccgcaagaa
     2881 aactatcccg accgccttac tgccgcctgt tttgaccgct gggatctgcc attgtcagac
     2941 atgtataccc cgtacgtctt cccgagcgaa aacggtctgc gctgcgggac gcgcgaattg
     3001 aattatggcc cacaccagtg gcgcggcgac ttccagttca acatcagccg ctacagtcaa
     3061 cagcaactga tggaaaccag ccatcgccat ctgctgcacg cggaagaagg cacatggctg
     3121 aatatcgacg gtttccatat ggggattggt ggcgacgact cctggagccc gtcagtatcg
     3181 gcggaattcc aactgagcgc cggtcgctac cattaccaac ttgtctggtg tcaaaaataa
     3241 taggcgaaac aaatcatctg acaccaccac cgtctgatgg atcgttctca tctccgtctc
     3301 cacattatta tccgacgact acatcgacac cgaatcgaat ggaaacaagt ccggagtaca
     3361 tgtttaacca tgaaatggtg ggtagatgat tattaaaatg tttaagaaaa ttaaataatt
     3421 tgttttaggc accaccggtc aatgcgatgt ggtatactac accacctcct tatcaagatc
     3481 caaactatcg tcatgtgcct ccaaatactg catttcaaaa tgcagagcaa atgaatggct
     3541 ccttctactg ttaatctatt taattcatta atttttcatt tattgactgt atcccggatg
     3601 tttcttgtcc tcccaacata tctcctaact gctcggttca ttttaaatat gctcatctca
     3661 ctacatcacc cagacactgg tccccacaga gttttttgta tactatttcg ggtcattttt
     3721 cttattctag actaatattg taagctataa gttgtagaat aattattgat ccaaatcaga
     3781 ttaagagtat aagctttgtt ttttctcctt ttctttataa cttgttacaa tttttgaaat
     3841 tccctttttt gacaggcttt tattacactg taactgtgtt tcttatcttg caaacattta
     3901 atgaattgta attctttagt atcttgaggg ctttttgttt ttcgaattat tgaagctcaa
     3961 agttccagtt ttactacgat ccagcgaatt ctcctcattt cgatccgatg caattgactt
     4021 cagatcaata ttggttgcct gaaagaaata attgtgagca tttttgtcaa aaaacagaga
     4081 actcaccatt ctcgaggctc ccgttccagg agcagtactt ggtgatggac acgtagattg
     4141 attaaaccaa accaaaggtt ctttcagagt caacttacag cctcgagcgt agtccgtgat
     4201 agcttctcgc agaacactga aaattggaaa tttattggaa taaaaacttt ttctgcactt
     4261 tatagaataa aaaaatcatg aatttacccg aatttaacct ccgaatcgta gaccaaattg
     4321 tccaagtaga tggaaatcac cttgaacatc ggatgttttt catatgctga aaataaatta
     4381 atgaatttat gtaatttttt aaataattac ttttcaattt ggtgaacaat tcctgcttct
     4441 ttgcataggc atctggacga gtgagtcctt tccaatcaat caatgtggtg tcgacctcga
     4501 gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaattccg agcttggcgt
     4561 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4621 taggagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagg taactcacat
     4681 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4741 aatgaatcgg ccaacgccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg
     4801 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4861 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4921 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     4981 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg
     5041 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5101 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5161 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5221 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5281 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5341 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5401 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5461 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5521 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5581 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5641 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5701 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5761 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5821 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5881 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5941 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6001 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6061 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6121 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6181 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6241 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6301 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6361 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6421 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6481 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6541 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6601 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
     6661 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6721 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6781 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6841 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6901 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6961 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7021 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7081 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7141 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     7201 gtggaattgt gagcggataa caatttcaca caggaaacag ct
//
