LOCUS       sequence-48-expo        8255 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 8255)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 8255)
  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: pPD20_97
FEATURES             Location/Qualifiers
     source          1..8255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1343..1363
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1451..4498
                     /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(1453..1475)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5829..5851)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5951..5970
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6048..6066)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6134..6238
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6239..6307,6308..7099)
                     /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(6457..6476)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7270..7858
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7759..7778
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8012..8029
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8146..8167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8182..8187,8188..8205,8206..8212)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8220..8236
                     /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     8225..8247
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8244..8255,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8244..8255,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccagctt
       61 gcatgcctgc aggtcgaggc atttgaattg ggggtggtgg acagtaactg tctgtaataa
      121 taattactcc tgaccaggtt gcaattcgag ttttgataag cataattata ccttgtacat
      181 tgtgggtttt gtgctgtgga cgttttattg tgacatcccc ataagctaca agaacaaaaa
      241 tgaaatgtaa aagtattgaa aacgtcgtac attttatatc tgagtagtat cctttgcttt
      301 aaatgtccat aaaactaatt ttataatcaa taaaacaacg tttgtaaatc aactgagttt
      361 acaagtagag acattgaggg atactttcac tatgctaaag tgataatcga ccaaataata
      421 acttcacttt ggtatttatt cctgtcttat aaatgttatg tatgaattaa attcatatgc
      481 atatggctca ctctgacaat ttttaataat cttccagatc aatattgact accgatgcgg
      541 gtggtctttt gctttgaatt ctgctgaact ttacaccccg aacagcaatg tgtgcttcag
      601 cctaaaaaaa agtaagtgtg ttaatcagtg cccccgattc ttcatttttt gcccctctct
      661 cccgtttcgt cggcaaaaga agagaaaata aagataagtc tcaagatagg ttggtaatcg
      721 ctaaagtggt tgtgtggata agagtagcaa aatggcagga agagcacttt gcgcgcacac
      781 actgtactca ttgttctgga taaaattctc tcgttgtttg ccgtcggatg tctgcctctc
      841 tgcattgagc cggcttcttc actatcttta gttaacctaa aatgccgttt cttttctcgt
      901 atccccacta tcccgttgag gttctctgct ctcttcgctc cctaccgcca gcgagcaact
      961 atccgtgggg gcgccttgct cggaagatgg gggggaagaa agaagatttt tgctatttgc
     1021 acttgagaaa gagacttttc ctgcgtcgat ggttagagaa cagtgtgcag acacttttca
     1081 gctacctaga attacaattg gatatccccg cctcccaatc cacccaccca gggaaaaaga
     1141 agggctcgcc gaaaatcaaa gttatctcca ggctcgcgca tcccaccgag cggttgactt
     1201 ctctccacca cttttcattt taaccctcgg ggtacgggat tggccaaagg acccaaaggt
     1261 atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt ggagggtacc
     1321 gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag gtaccggtgg gtgaagacca
     1381 gaaacagcac ctcgaactga gccgcgatat tgcccagcgt ttcaacgcgc tgtatggcga
     1441 gatcgatccc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     1501 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     1561 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc
     1621 accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt
     1681 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta
     1741 tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct
     1801 cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg
     1861 cgttaactcg gcgtttcatc tgtggtgcaa cgggcgctgg gtcggttacg gccaggacag
     1921 tcgtttgccg tctgaatttg acctgagcgc atttttacgc gccggagaaa accgcctcgc
     1981 ggtgatggtg ctgcgctgga gtgacggcag ttatctggaa gatcaggata tgtggcggat
     2041 gagcggcatt ttccgtgacg tctcgttgct gcataaaccg actacacaaa tcagcgattt
     2101 ccatgttgcc actcgcttta atgatgattt cagccgcgct gtactggagg ctgaagttca
     2161 gatgtgcggc gagttgcgtg actacctacg ggtaacagtt tctttatggc agggtgaaac
     2221 gcaggtcgcc agcggcaccg cgcctttcgg cggtgaaatt atcgatgagc gtggtggtta
     2281 tgccgatcgc gtcacactac gtctgaacgt cgaaaacccg aaactgtgga gcgccgaaat
     2341 cccgaatctc tatcgtgcgg tggttgaact gcacaccgcc gacggcacgc tgattgaagc
     2401 agaagcctgc gatgtcggtt tccgcgaggt gcggattgaa aatggtctgc tgctgctgaa
     2461 cggcaagccg ttgctgattc gaggcgttaa ccgtcacgag catcatcctc tgcatggtca
     2521 ggtcatggat gagcagacga tggtgcagga tatcctgctg atgaagcaga acaactttaa
     2581 cgccgtgcgc tgttcgcatt atccgaacca tccgctgtgg tacacgctgt gcgaccgcta
     2641 cggcctgtat gtggtggatg aagccaatat tgaaacccac ggcatggtgc caatgaatcg
     2701 tctgaccgat gatccgcgct ggctaccggc gatgagcgaa cgcgtaacgc gaatggtgca
     2761 gcgcgatcgt aatcacccga gtgtgatcat ctggtcgctg gggaatgaat caggccacgg
     2821 cgctaatcac gacgcgctgt atcgctggat caaatctgtc gatccttccc gcccggtgca
     2881 gtatgaaggc ggcggagccg acaccacggc caccgatatt atttgcccga tgtacgcgcg
     2941 cgtggatgaa gaccagccct tcccggctgt gccgaaatgg tccatcaaaa aatggctttc
     3001 gctacctgga gagacgcgcc cgctgatcct ttgcgaatac gcccacgcga tgggtaacag
     3061 tcttggcggt ttcgctaaat actggcaggc gtttcgtcag tatccccgtt tacagggcgg
     3121 cttcgtctgg gactgggtgg atcagtcgct gattaaatat gatgaaaacg gcaacccgtg
     3181 gtcggcttac ggcggtgatt ttggcgatac gccgaacgat cgccagttct gtatgaacgg
     3241 tctggtcttt gccgaccgca cgccgcatcc agcgctgacg gaagcaaaac accagcagca
     3301 gtttttccag ttccgtttat ccgggcaaac catcgaagtg accagcgaat acctgttccg
     3361 tcatagcgat aacgagctcc tgcactggat ggtggcgctg gatggtaagc cgctggcaag
     3421 cggtgaagtg cctctggatg tcgctccaca aggtaaacag ttgattgaac tgcctgaact
     3481 accgcagccg gagagcgccg ggcaactctg gctcacagta cgcgtagtgc aaccgaacgc
     3541 gaccgcatgg tcagaagccg ggcacatcag cgcctggcag cagtggcgtc tggcggaaaa
     3601 cctcagtgtg acgctccccg ccgcgtccca cgccatcccg catctgacca ccagcgaaat
     3661 ggatttttgc atcgagctgg gtaataagcg ttggcaattt aaccgccagt caggctttct
     3721 ttcacagatg tggattggcg ataaaaaaca actgctgacg ccgctgcgcg atcagttcac
     3781 ccgtgcaccg ctggataacg acattggcgt aagtgaagcg acccgcattg accctaacgc
     3841 ctgggtcgaa cgctggaagg cggcgggcca ttaccaggcc gaagcagcgt tgttgcagtg
     3901 cacggcagat acacttgctg atgcggtgct gattacgacc gctcacgcgt ggcagcatca
     3961 ggggaaaacc ttatttatca gccggaaaac ctaccggatt gatggtagtg gtcaaatggc
     4021 gattaccgtt gatgttgaag tggcgagcga tacaccgcat ccggcgcgga ttggcctgaa
     4081 ctgccagctg gcgcaggtag cagagcgggt aaactggctc ggattagggc cgcaagaaaa
     4141 ctatcccgac cgccttactg ccgcctgttt tgaccgctgg gatctgccat tgtcagacat
     4201 gtataccccg tacgtcttcc cgagcgaaaa cggtctgcgc tgcgggacgc gcgaattgaa
     4261 ttatggccca caccagtggc gcggcgactt ccagttcaac atcagccgct acagtcaaca
     4321 gcaactgatg gaaaccagcc atcgccatct gctgcacgcg gaagaaggca catggctgaa
     4381 tatcgacggt ttccatatgg ggattggtgg cgacgactcc tggagcccgt cagtatcggc
     4441 ggaattccaa ctgagcgccg gtcgctacca ttaccaactt gtctggtgtc aaaaataata
     4501 ggggccgctg tcatcagatc gccatctcgc gcccgtgcct ctgacttcta agtccaatta
     4561 ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta
     4621 tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg
     4681 aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct
     4741 ccctcccccc attaataata attctatccc aaaatctaca caatgttctg tgtacacttc
     4801 ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa
     4861 taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg
     4921 ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt
     4981 tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc
     5041 ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg
     5101 agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta
     5161 gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat
     5221 acattcccaa tataccaaac ataactgttt aaaattaaac atttttctaa attttatatg
     5281 atttctttta aatttgcaaa aattacttaa atttgaattc ccgcgcaaat gagtgacttc
     5341 attttctgca ttattgtgtt ttccggctat attaataggt atttgtttgt gtttttcttt
     5401 attttatgat tcgaactcca atttgtaaat tttcgaacat atttccctaa agaaaaaata
     5461 tgattaatct ggaaaaattg gaaaattatt tttcaaataa aaaacaaaga aaaaaatgaa
     5521 gaaaaaccta ttagtttggc cataaaacgc aaaaatgtcg aaaatgacgt cactcatctg
     5581 cgcgggaaat caagaataat tcggcctttt ttattttttt ggaaaatcgt aaaacattta
     5641 gaaaaatttt ttaatagtta tatgggactt attctgcatt tagggcaaaa gccagagact
     5701 actccaccgt taacatgaat tatgaatatt attgcgacaa gacccaaaca ttgataaacc
     5761 caaatctagc ctactagtcg gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac
     5821 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5881 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     5941 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     6001 ccgcacagat gcgtaaggag aaaataccgc atcaggcggc cttaagggcc tcgtgatacg
     6061 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     6121 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6181 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6241 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6301 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6361 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6421 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6481 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6541 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6601 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6661 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6721 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6781 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6841 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6901 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6961 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7021 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7081 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7141 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7201 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7261 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7321 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7381 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     7441 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7501 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7561 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7621 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     7681 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7741 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7801 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7861 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7921 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7981 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     8041 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     8101 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     8161 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     8221 tgtgagcgga taacaatttc acacaggaaa cagct
//
