LOCUS       sequence-68-expo        6554 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 6554)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6554)
  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: pPD81_34
FEATURES             Location/Qualifiers
     source          1..6554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             161..181
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             269..3316
                     /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(271..293)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4128..4150)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4250..4269
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4347..4365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4433..4537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4538..4606,4607..5398)
                     /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(4756..4775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5569..6157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6058..6077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6311..6328
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6445..6466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6481..6486,6487..6504,6505..6511)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6519..6535
                     /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     6524..6546
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6543..6554,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6543..6554,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctagaggatc cccgggattg
       61 gccaaaggac ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga
      121 ggacccttgg agggtaccga gctcagaaaa aatgactgct ccaaagaaga agcgtaaggt
      181 accggtgggt gaagaccaga aacagcacct cgaactgagc cgcgatattg cccagcgttt
      241 caacgcgctg tatggcgaga tcgatcccgt cgttttacaa cgtcgtgact gggaaaaccc
      301 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
      361 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
      421 ctttgcctgg tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc
      481 tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat
      541 ctacaccaac gtgacctatc ccattacggt caatccgccg tttgttccca cggagaatcc
      601 gacgggttgt tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac
      661 gcgaattatt tttgatggcg ttaactcggc gtttcatctg tggtgcaacg ggcgctgggt
      721 cggttacggc caggacagtc gtttgccgtc tgaatttgac ctgagcgcat ttttacgcgc
      781 cggagaaaac cgcctcgcgg tgatggtgct gcgctggagt gacggcagtt atctggaaga
      841 tcaggatatg tggcggatga gcggcatttt ccgtgacgtc tcgttgctgc ataaaccgac
      901 tacacaaatc agcgatttcc atgttgccac tcgctttaat gatgatttca gccgcgctgt
      961 actggaggct gaagttcaga tgtgcggcga gttgcgtgac tacctacggg taacagtttc
     1021 tttatggcag ggtgaaacgc aggtcgccag cggcaccgcg cctttcggcg gtgaaattat
     1081 cgatgagcgt ggtggttatg ccgatcgcgt cacactacgt ctgaacgtcg aaaacccgaa
     1141 actgtggagc gccgaaatcc cgaatctcta tcgtgcggtg gttgaactgc acaccgccga
     1201 cggcacgctg attgaagcag aagcctgcga tgtcggtttc cgcgaggtgc ggattgaaaa
     1261 tggtctgctg ctgctgaacg gcaagccgtt gctgattcga ggcgttaacc gtcacgagca
     1321 tcatcctctg catggtcagg tcatggatga gcagacgatg gtgcaggata tcctgctgat
     1381 gaagcagaac aactttaacg ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta
     1441 cacgctgtgc gaccgctacg gcctgtatgt ggtggatgaa gccaatattg aaacccacgg
     1501 catggtgcca atgaatcgtc tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg
     1561 cgtaacgcga atggtgcagc gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg
     1621 gaatgaatca ggccacggcg ctaatcacga cgcgctgtat cgctggatca aatctgtcga
     1681 tccttcccgc ccggtgcagt atgaaggcgg cggagccgac accacggcca ccgatattat
     1741 ttgcccgatg tacgcgcgcg tggatgaaga ccagcccttc ccggctgtgc cgaaatggtc
     1801 catcaaaaaa tggctttcgc tacctggaga gacgcgcccg ctgatccttt gcgaatacgc
     1861 ccacgcgatg ggtaacagtc ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta
     1921 tccccgttta cagggcggct tcgtctggga ctgggtggat cagtcgctga ttaaatatga
     1981 tgaaaacggc aacccgtggt cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg
     2041 ccagttctgt atgaacggtc tggtctttgc cgaccgcacg ccgcatccag cgctgacgga
     2101 agcaaaacac cagcagcagt ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac
     2161 cagcgaatac ctgttccgtc atagcgataa cgagctcctg cactggatgg tggcgctgga
     2221 tggtaagccg ctggcaagcg gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt
     2281 gattgaactg cctgaactac cgcagccgga gagcgccggg caactctggc tcacagtacg
     2341 cgtagtgcaa ccgaacgcga ccgcatggtc agaagccggg cacatcagcg cctggcagca
     2401 gtggcgtctg gcggaaaacc tcagtgtgac gctccccgcc gcgtcccacg ccatcccgca
     2461 tctgaccacc agcgaaatgg atttttgcat cgagctgggt aataagcgtt ggcaatttaa
     2521 ccgccagtca ggctttcttt cacagatgtg gattggcgat aaaaaacaac tgctgacgcc
     2581 gctgcgcgat cagttcaccc gtgcaccgct ggataacgac attggcgtaa gtgaagcgac
     2641 ccgcattgac cctaacgcct gggtcgaacg ctggaaggcg gcgggccatt accaggccga
     2701 agcagcgttg ttgcagtgca cggcagatac acttgctgat gcggtgctga ttacgaccgc
     2761 tcacgcgtgg cagcatcagg ggaaaacctt atttatcagc cggaaaacct accggattga
     2821 tggtagtggt caaatggcga ttaccgttga tgttgaagtg gcgagcgata caccgcatcc
     2881 ggcgcggatt ggcctgaact gccagctggc gcaggtagca gagcgggtaa actggctcgg
     2941 attagggccg caagaaaact atcccgaccg ccttactgcc gcctgttttg accgctggga
     3001 tctgccattg tcagacatgt ataccccgta cgtcttcccg agcgaaaacg gtctgcgctg
     3061 cgggacgcgc gaattgaatt atggcccaca ccagtggcgc ggcgacttcc agttcaacat
     3121 cagccgctac agtcaacagc aactgatgga aaccagccat cgccatctgc tgcacgcgga
     3181 agaaggcaca tggctgaata tcgacggttt ccatatgggg attggtggcg acgactcctg
     3241 gagcccgtca gtatcggcgg aattccaact gagcgccggt cgctaccatt accaacttgt
     3301 ctggtgtcaa aaataatagg ggccgctgtc atcagatcgc catctcgcgc ccgtgcctct
     3361 gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct gtgctcccac
     3421 cccctatttt tgttattatc aaaaaacttc tcttaatttc tttgtttttt agcttctttt
     3481 aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc gtaataaaaa
     3541 gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa aatctacaca
     3601 atgttctgtg tacacttctt atgtttttta cttctgataa atttttttga aacatcatag
     3661 aaaaaaccgc acacaaaata ccttatcata tgttacgttt cagtttatga ccgcaatttt
     3721 tatttcttcg cacgtctggg cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt
     3781 tggagtattt ttggaatttt tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt
     3841 tgctttttgg ggtttcccct attgtttgtc aagatttcga ggacggcgtt tttcttgcta
     3901 aaatcacaag tattgatgag cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg
     3961 ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg agtagtgtct atggggtttt
     4021 tgccttaaat gacagaatac attcccaata taccaaacat aactgtttcc tactagtcgg
     4081 ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4141 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4201 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4261 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4321 aaataccgca tcaggcggcc ttaagggcct cgtgatacgc ctatttttat aggttaatgt
     4381 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4441 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4501 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4561 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4621 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4681 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4741 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4801 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4861 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4921 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4981 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5041 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5101 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5161 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5221 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5281 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5341 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5401 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5461 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5521 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5581 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5641 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5701 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5761 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5821 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5881 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5941 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6001 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6061 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6121 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6181 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6241 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6301 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6361 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6421 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6481 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6541 cacaggaaac agct
//
