LOCUS       sequence-65-expo        8217 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 8217)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 8217)
  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: pPD69_39
FEATURES             Location/Qualifiers
     source          1..8217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1772..1792
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1880..4927
                     /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(1882..1904)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5791..5813)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5913..5932
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6010..6028)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6096..6200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6201..6269,6270..7061)
                     /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(6419..6438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7232..7820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7721..7740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7974..7991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8108..8129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8144..8149,8150..8167,8168..8174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8182..8198
                     /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     8187..8209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8206..8217,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8206..8217,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggcctt ggtcgagtat gagaaacaac
       61 attaacaatg ttttgggaga aacatgtagt tttgtacgtc cggtccagtg agagttttca
      121 acttattttg gaagaaattt ttttttgaca aaattttgca atttcccacg cagaaatacg
      181 gtatgccccg tatctcgcca cgaacataat attttaaatt gacggaaagg tgtgcgcctt
      241 taagaagcac agtaaccatt ttccggtttt ttttttgaaa ttgcctttga tttttacttt
      301 tttctcccaa aaacgagaaa tttaatataa tccatcaaaa tctttgtgaa cgagagaaat
      361 atagtgaaaa actcaaagaa aattgcgaaa tggattactg tactccataa aggcgcacat
      421 cttttcgaaa aatcgtgaaa aattgcatct gaccaacagt tttgcgtgga atcttcctag
      481 attcaaattt cctatgagtc ataaaactat cattttatcg gacaataact acacgtgtac
      541 cttgtttcca tatgttctac caatataatt tgtgttcata tgtggccagg tacgggaaat
      601 tagaacacgg actgatttag gaaggttcaa attcgagaac aatggacgac ttgctgcacg
      661 aaaacgtgga ttattctgaa accgcctatg tgatcactat cgatttagca cttcggtttt
      721 ctttttccgt tttcctttat attccgtatt gtatgccgac aataacgctt tttctttgta
      781 taaacatttc cagaagcaga gcaaatttgt ttattccaga attgactgaa tattgaaata
      841 atgtttacag acatactgta gatttttttc tattcaaaca tgaatacagt ttttcgtatt
      901 tttaatcttc agataagttt tgtgtgtgtt tttgagaata atatttaaaa aagataagca
      961 aatatcagaa ctacggtaag cgaaggggta cggtcgtcaa ttgcggactt ccgagctgtg
     1021 cacggcgcca aaatttattt tgagttttaa aatgattttc tttgtttttt tttctttttt
     1081 tgacacgata agaacatttt tctgtatttc taatatttta aatgagatcg taataagttt
     1141 atttagctcc atcacattat tttcggaata ttcatcgctt gaaaacggcg taaaataggt
     1201 tccaagtaaa tcattgcaaa caaatctccg tttatcgttc taacaaagtt tttcgttcac
     1261 tttcttatgt aaatcgtttt ccccacttct tcttcacggg aatgatcaat tgaaaggcca
     1321 taaattggtt gttttctcca catttcgcaa ccaacatgca gctgtcggat aggcaataaa
     1381 ataatgtaat caatgagttt tccgatttat cctaacagtt ctttttctag tgttcctatt
     1441 cgaaaaaagt actgttatcc ttctttttgg accgcccatc tcaggtgtca agacaatcgc
     1501 aacacactca gaaaatgggc ggagtatgcc acccggcctt cattcgtcgc tccttgtctt
     1561 ctccattttc gtttttttcc tgttctccgt tctttttttg tacttttgtg ataatgctga
     1621 agtgttcttt tttttcgctt aaccgcatct cttgatttaa ttcatatttt ctttcagatt
     1681 catatacaat aacaccaaga gctgctctaa caactaataa taacagaatc tagaggatcc
     1741 ccgggtaccg agctcagaaa aaatgactgc tccaaagaag aagcgtaagg taccggtggg
     1801 tgaagaccag aaacagcacc tcgaactgag ccgcgatatt gcccagcgtt tcaacgcgct
     1861 gtatggcgag atcgatcccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     1921 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     1981 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg
     2041 gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga
     2101 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa
     2161 cgtgacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg
     2221 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat
     2281 ttttgatggc gttaactcgg cgtttcatct gtggtgcaac gggcgctggg tcggttacgg
     2341 ccaggacagt cgtttgccgt ctgaatttga cctgagcgca tttttacgcg ccggagaaaa
     2401 ccgcctcgcg gtgatggtgc tgcgctggag tgacggcagt tatctggaag atcaggatat
     2461 gtggcggatg agcggcattt tccgtgacgt ctcgttgctg cataaaccga ctacacaaat
     2521 cagcgatttc catgttgcca ctcgctttaa tgatgatttc agccgcgctg tactggaggc
     2581 tgaagttcag atgtgcggcg agttgcgtga ctacctacgg gtaacagttt ctttatggca
     2641 gggtgaaacg caggtcgcca gcggcaccgc gcctttcggc ggtgaaatta tcgatgagcg
     2701 tggtggttat gccgatcgcg tcacactacg tctgaacgtc gaaaacccga aactgtggag
     2761 cgccgaaatc ccgaatctct atcgtgcggt ggttgaactg cacaccgccg acggcacgct
     2821 gattgaagca gaagcctgcg atgtcggttt ccgcgaggtg cggattgaaa atggtctgct
     2881 gctgctgaac ggcaagccgt tgctgattcg aggcgttaac cgtcacgagc atcatcctct
     2941 gcatggtcag gtcatggatg agcagacgat ggtgcaggat atcctgctga tgaagcagaa
     3001 caactttaac gccgtgcgct gttcgcatta tccgaaccat ccgctgtggt acacgctgtg
     3061 cgaccgctac ggcctgtatg tggtggatga agccaatatt gaaacccacg gcatggtgcc
     3121 aatgaatcgt ctgaccgatg atccgcgctg gctaccggcg atgagcgaac gcgtaacgcg
     3181 aatggtgcag cgcgatcgta atcacccgag tgtgatcatc tggtcgctgg ggaatgaatc
     3241 aggccacggc gctaatcacg acgcgctgta tcgctggatc aaatctgtcg atccttcccg
     3301 cccggtgcag tatgaaggcg gcggagccga caccacggcc accgatatta tttgcccgat
     3361 gtacgcgcgc gtggatgaag accagccctt cccggctgtg ccgaaatggt ccatcaaaaa
     3421 atggctttcg ctacctggag agacgcgccc gctgatcctt tgcgaatacg cccacgcgat
     3481 gggtaacagt cttggcggtt tcgctaaata ctggcaggcg tttcgtcagt atccccgttt
     3541 acagggcggc ttcgtctggg actgggtgga tcagtcgctg attaaatatg atgaaaacgg
     3601 caacccgtgg tcggcttacg gcggtgattt tggcgatacg ccgaacgatc gccagttctg
     3661 tatgaacggt ctggtctttg ccgaccgcac gccgcatcca gcgctgacgg aagcaaaaca
     3721 ccagcagcag tttttccagt tccgtttatc cgggcaaacc atcgaagtga ccagcgaata
     3781 cctgttccgt catagcgata acgagctcct gcactggatg gtggcgctgg atggtaagcc
     3841 gctggcaagc ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt tgattgaact
     3901 gcctgaacta ccgcagccgg agagcgccgg gcaactctgg ctcacagtac gcgtagtgca
     3961 accgaacgcg accgcatggt cagaagccgg gcacatcagc gcctggcagc agtggcgtct
     4021 ggcggaaaac ctcagtgtga cgctccccgc cgcgtcccac gccatcccgc atctgaccac
     4081 cagcgaaatg gatttttgca tcgagctggg taataagcgt tggcaattta accgccagtc
     4141 aggctttctt tcacagatgt ggattggcga taaaaaacaa ctgctgacgc cgctgcgcga
     4201 tcagttcacc cgtgcaccgc tggataacga cattggcgta agtgaagcga cccgcattga
     4261 ccctaacgcc tgggtcgaac gctggaaggc ggcgggccat taccaggccg aagcagcgtt
     4321 gttgcagtgc acggcagata cacttgctga tgcggtgctg attacgaccg ctcacgcgtg
     4381 gcagcatcag gggaaaacct tatttatcag ccggaaaacc taccggattg atggtagtgg
     4441 tcaaatggcg attaccgttg atgttgaagt ggcgagcgat acaccgcatc cggcgcggat
     4501 tggcctgaac tgccagctgg cgcaggtagc agagcgggta aactggctcg gattagggcc
     4561 gcaagaaaac tatcccgacc gccttactgc cgcctgtttt gaccgctggg atctgccatt
     4621 gtcagacatg tataccccgt acgtcttccc gagcgaaaac ggtctgcgct gcgggacgcg
     4681 cgaattgaat tatggcccac accagtggcg cggcgacttc cagttcaaca tcagccgcta
     4741 cagtcaacag caactgatgg aaaccagcca tcgccatctg ctgcacgcgg aagaaggcac
     4801 atggctgaat atcgacggtt tccatatggg gattggtggc gacgactcct ggagcccgtc
     4861 agtatcggcg gaattccaac tgagcgccgg tcgctaccat taccaacttg tctggtgtca
     4921 aaaataatag gcctactagc gtcgacggta ccatgcatgg tattgatatc tgagctccgc
     4981 atcggccgct gtcatcagat cgccatctcg cgcccgtgcc tctgacttct aagtccaatt
     5041 actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt
     5101 atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat
     5161 gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc
     5221 tccctccccc cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt
     5281 cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa
     5341 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct
     5401 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat
     5461 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc
     5521 cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     5581 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     5641 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     5701 tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg
     5761 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5821 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5881 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5941 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6001 gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6061 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     8161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagct
//
