LOCUS       sequence-96-expo        7101 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 7101)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7101)
  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: pPD95_21
FEATURES             Location/Qualifiers
     source          1..7101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             527..547
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             635..3682
                     /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(637..659)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4541..4563)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4663..4682
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4760..4778)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4846..4950
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4951..5019,5020..5811)
                     /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(5169..5188)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5982..6570
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6471..6490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6724..6741
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6858..6879
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6894..6899,6900..6917,6918..6924)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6932..6948
                     /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     6937..6959
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6956..6972
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6956..6972
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagaggat ccccggggat tggccaaagg
       61 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
      121 gcttggctag caaaaatgca taaggttttg ctggcactgt tctttatctt tctggcacca
      181 gcatccgcac tggcagtctc cgaaccggcc tgcaggatcg attttttgca aattacgagc
      241 gttgtagggg gcggagcgat aggtcctata ggttttggta tatcatcatt cattcattca
      301 ttggtacatt catttaccca ccttcctctt tctgagcttc tctggagttc tgtgcttcct
      361 ttttccctta tctttatact gtaattttta actttcaggc attgattgga tccccgggat
      421 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      481 gaggaccctt gcttggaggg taccgagctc agaaaaaatg actgctccaa agaagaagcg
      541 taaggtaccg gtgggtgaag accagaaaca gcacctcgaa ctgagccgcg atattgccca
      601 gcgtttcaac gcgctgtatg gcgagatcga tcccgtcgtt ttacaacgtc gtgactggga
      661 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
      721 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
      781 atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga
      841 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
      901 gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga
      961 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
     1021 ccagacgcga attatttttg atggcgttaa ctcggcgttt catctgtggt gcaacgggcg
     1081 ctgggtcggt tacggccagg acagtcgttt gccgtctgaa tttgacctga gcgcattttt
     1141 acgcgccgga gaaaaccgcc tcgcggtgat ggtgctgcgc tggagtgacg gcagttatct
     1201 ggaagatcag gatatgtggc ggatgagcgg cattttccgt gacgtctcgt tgctgcataa
     1261 accgactaca caaatcagcg atttccatgt tgccactcgc tttaatgatg atttcagccg
     1321 cgctgtactg gaggctgaag ttcagatgtg cggcgagttg cgtgactacc tacgggtaac
     1381 agtttcttta tggcagggtg aaacgcaggt cgccagcggc accgcgcctt tcggcggtga
     1441 aattatcgat gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa
     1501 cccgaaactg tggagcgccg aaatcccgaa tctctatcgt gcggtggttg aactgcacac
     1561 cgccgacggc acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat
     1621 tgaaaatggt ctgctgctgc tgaacggcaa gccgttgctg attcgaggcg ttaaccgtca
     1681 cgagcatcat cctctgcatg gtcaggtcat ggatgagcag acgatggtgc aggatatcct
     1741 gctgatgaag cagaacaact ttaacgccgt gcgctgttcg cattatccga accatccgct
     1801 gtggtacacg ctgtgcgacc gctacggcct gtatgtggtg gatgaagcca atattgaaac
     1861 ccacggcatg gtgccaatga atcgtctgac cgatgatccg cgctggctac cggcgatgag
     1921 cgaacgcgta acgcgaatgg tgcagcgcga tcgtaatcac ccgagtgtga tcatctggtc
     1981 gctggggaat gaatcaggcc acggcgctaa tcacgacgcg ctgtatcgct ggatcaaatc
     2041 tgtcgatcct tcccgcccgg tgcagtatga aggcggcgga gccgacacca cggccaccga
     2101 tattatttgc ccgatgtacg cgcgcgtgga tgaagaccag cccttcccgg ctgtgccgaa
     2161 atggtccatc aaaaaatggc tttcgctacc tggagagacg cgcccgctga tcctttgcga
     2221 atacgcccac gcgatgggta acagtcttgg cggtttcgct aaatactggc aggcgtttcg
     2281 tcagtatccc cgtttacagg gcggcttcgt ctgggactgg gtggatcagt cgctgattaa
     2341 atatgatgaa aacggcaacc cgtggtcggc ttacggcggt gattttggcg atacgccgaa
     2401 cgatcgccag ttctgtatga acggtctggt ctttgccgac cgcacgccgc atccagcgct
     2461 gacggaagca aaacaccagc agcagttttt ccagttccgt ttatccgggc aaaccatcga
     2521 agtgaccagc gaatacctgt tccgtcatag cgataacgag ctcctgcact ggatggtggc
     2581 gctggatggt aagccgctgg caagcggtga agtgcctctg gatgtcgctc cacaaggtaa
     2641 acagttgatt gaactgcctg aactaccgca gccggagagc gccgggcaac tctggctcac
     2701 agtacgcgta gtgcaaccga acgcgaccgc atggtcagaa gccgggcaca tcagcgcctg
     2761 gcagcagtgg cgtctggcgg aaaacctcag tgtgacgctc cccgccgcgt cccacgccat
     2821 cccgcatctg accaccagcg aaatggattt ttgcatcgag ctgggtaata agcgttggca
     2881 atttaaccgc cagtcaggct ttctttcaca gatgtggatt ggcgataaaa aacaactgct
     2941 gacgccgctg cgcgatcagt tcacccgtgc accgctggat aacgacattg gcgtaagtga
     3001 agcgacccgc attgacccta acgcctgggt cgaacgctgg aaggcggcgg gccattacca
     3061 ggccgaagca gcgttgttgc agtgcacggc agatacactt gctgatgcgg tgctgattac
     3121 gaccgctcac gcgtggcagc atcaggggaa aaccttattt atcagccgga aaacctaccg
     3181 gattgatggt agtggtcaaa tggcgattac cgttgatgtt gaagtggcga gcgatacacc
     3241 gcatccggcg cggattggcc tgaactgcca gctggcgcag gtagcagagc gggtaaactg
     3301 gctcggatta gggccgcaag aaaactatcc cgaccgcctt actgccgcct gttttgaccg
     3361 ctgggatctg ccattgtcag acatgtatac cccgtacgtc ttcccgagcg aaaacggtct
     3421 gcgctgcggg acgcgcgaat tgaattatgg cccacaccag tggcgcggcg acttccagtt
     3481 caacatcagc cgctacagtc aacagcaact gatggaaacc agccatcgcc atctgctgca
     3541 cgcggaagaa ggcacatggc tgaatatcga cggtttccat atggggattg gtggcgacga
     3601 ctcctggagc ccgtcagtat cggcggaatt ccaactgagc gccggtcgct accattacca
     3661 acttgtctgg tgtcaaaaat aataggggcc gctgtcatca gagtaagttt aaactgagtt
     3721 ctactaacta acgagtaata tttaaatttt cagcatctcg cgcccgtgcc tctgacttct
     3781 aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat
     3841 ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac
     3901 ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa
     3961 aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac acaatgttct
     4021 gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac
     4081 cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct
     4141 tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta
     4201 tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt
     4261 tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac
     4321 aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg
     4381 gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta
     4441 aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg
     4501 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4561 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4621 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4681 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4741 gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4801 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4861 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4921 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4981 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5041 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5101 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5161 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5221 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5281 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5341 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5401 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5461 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5521 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5581 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5641 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5701 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5761 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5821 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5881 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5941 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6001 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6061 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6121 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6361 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6601 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6661 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6721 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6781 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6841 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6901 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6961 aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta ctaactaact
     7021 attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac gatggatacg
     7081 ctaacaactt ggaaatgaaa t
//
