LOCUS       sequence-99-expo        7228 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 7228)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7228)
  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_57
FEATURES             Location/Qualifiers
     source          1..7228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             145..165
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(255..277)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4668..4690)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4790..4809
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4887..4905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4973..5077
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5078..5146,5147..5938)
                     /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(5296..5315)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6109..6697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6598..6617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6851..6868
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6985..7006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7021..7026,7027..7044,7045..7051)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7059..7075
                     /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     7064..7086
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7083..7099
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7083..7099
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gattggccaa aggacccaaa
       61 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttgagggta
      121 ccgagctcag aaaaaatgac tgctccaaag aagaagcgta aggtaccggt gggtgaagac
      181 cagaaacagc acctcgaact gagccgcgat attgcccagc gtttcaacgc gctgtatggc
      241 gagatcgatc ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
      301 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
      361 gatcgccctt cccaacagtt gcgaaggtaa gtttaaacag atccatacta actaacttgt
      421 tctgacataa ttttcagctt gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
      481 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
      541 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
      601 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
      661 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttaac
      721 tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt acggccaggt aagtttaatt
      781 aagttgatac taactaacaa gatctgatta attttcagga cagtcgtttg ccgtctgaat
      841 ttgacctgag cgcattttta cgcgccggag aaaaccgcct cgcggtgatg gtgctgcgct
      901 ggagtgacgg cagttatctg gaagatcagg atatgtggcg gatgagcggc attttccgtg
      961 acgtctcgtt gctgcataaa ccgactacac aaatcagcga tttccatgtt gccactcgct
     1021 ttaatgatga tttctcccgc gctgtactgg aggctgaagt ccaggtaagt ttaaacagga
     1081 tcttactaac taacatgcta acactgaatt ttcagatgtg cggcgagttg cgtgactacc
     1141 tacgggtaac agtttcttta tggcagggtg aaacgcaggt cgccagcggc accgcgcctt
     1201 tcggcggtga aattatcgat gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga
     1261 acgtcgaaaa cccgaaactg tggagcgccg aaatcccgaa tctctatcgt gcggtggttg
     1321 aactgcacac cgccgacggc acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg
     1381 aggtgcggat tgaaaatggt ctgctgctgc tgaacggcaa gccgttgctg attcgaggcg
     1441 ttaaccgtca cgagcatcat cctctgcatg gtcaggtcat ggatgagcag acgatggtgc
     1501 aggatgtaag tttaaactat tcgttactaa ctaactttaa acatttaaat tttcagatcc
     1561 tgctgatgaa gcagaacaac tttaacgccg tgcgctgttc gcattatccg aaccatccgc
     1621 tgtggtacac gctgtgcgac cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa
     1681 cccacggcat ggtgccaatg aatcgtctga ccgatgatcc gcgctggcta ccggcgatga
     1741 gcgaacgcgt aacgcgaatg gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt
     1801 cgctggggaa tggtaagttt aaacagttga atactaacta acggagatct ttgaaatttt
     1861 cagaatcagg ccacggcgct aatcacgacg cgctgtatcg ctggatcaaa tctgtcgatc
     1921 cttcccgccc ggtgcagtat gaaggcggcg gagccgacac cacggccacc gatattattt
     1981 gcccgatgta cgcgcgcgtg gatgaagacc agcccttccc ggctgtgccg aaatggtcca
     2041 tcaaaaaatg gctttcgcta cctggagaga cgcgcccgct gattctttgc gaggtaagtt
     2101 taaacagaac tctactaact aacacattag atcctaattt tcagtacgct cacgcgatgg
     2161 gcaacagtct tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat ccccgtttac
     2221 agggcggctt cgtctgggac tgggtggatc agtcgctgat taaatatgat gaaaacggca
     2281 acccgtggtc ggcttacggc ggtgattttg gcgatacgcc gaacgatcgc cagttctgta
     2341 tgaacggtct ggtctttgcc gaccgcacgc cgcatccagc gtaagtttaa acaataacct
     2401 actaactaac gtagataatt taaattttca ggctgacgga agcaaaacac cagcagcagt
     2461 ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac ctgttccgtc
     2521 atagcgataa cgagctcctg cactggatgg tggcgctgga tggtaagccg ctggcaagcg
     2581 gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt gattgaactg cctgaactac
     2641 cgcagccgga gagcgccggg caactctggc tcacagtacg cgtagtgcaa ccgaacgcga
     2701 ccgcatggtc agaagccggg cacatcagcg catggcagca gtggaggtaa gtttaaacaa
     2761 gatcctacta actaactcta cattgatgaa ttttcagact ggcggaaaac ctcagtgtga
     2821 cgctccccgc cgcgtcccac gccatcccgc atctgaccac cagcgaaatg gatttttgca
     2881 tcgagctggg taataagcgt tggcaattta accgccagtc aggctttctt tcacagatgt
     2941 ggattggcga taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc cgtgcaccgc
     3001 tggataacga cattggcgta agtgaagcga cccgcattga ccctaacgcc tgggtcggta
     3061 agtttaaaca aagttgtact aactaacgaa gatcttgata attttcagaa cgctggaagg
     3121 cggcgggcca ttaccaggcc gaagcagcgt tgttgcagtg cacggcagat acacttgctg
     3181 atgcggtgct gattacgacc gctcacgcgt ggcagcatca ggggaaaacc ttatttatca
     3241 gccggaaaac ctaccggatt gatggtagtg gtcaaatggc gattaccgtt gatgttgaag
     3301 tggcgagcga tacaccgcat ccggcgcgga ttggcctgaa ctgccagctg gcgcaggtag
     3361 cagagcgggt aaactggctc ggattagggc cgcaagaaaa ctatcccgac cgccttactg
     3421 ccgcctgttt tgaccgctgg gatctgccat tgtcagacat gtagtaagtt taaacttgat
     3481 agtactaact aacatgtttc atttaaattt tcagtacccc gtacgtcttc ccgagcgaaa
     3541 acggtctgcg ctgcgggacg cgcgaattga attatggccc acaccagtgg cgcggcgact
     3601 tccagttcaa catcagccgc tacagtcaac agcaactgat ggaaaccagc catcgccatc
     3661 tgctgcacgc ggaagaaggc acatggctga atatcgacgg tttccatatg gggattggtg
     3721 gcgacgactc ctggagcccg tcagtatcgg cggaattcca actgagcgcc ggtcgctacc
     3781 attaccaact tgtctggtgt caaaaataat aggggccgct gtcatcagag taagtttaaa
     3841 ctgagttcta ctaactaacg agtaatattt aaattttcag catctcgcgc ccgtgcctct
     3901 gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct gtgctcccac
     3961 cccctatttt tgttattatc aaaaaacttc tcttaatttc tttgtttttt agcttctttt
     4021 aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc gtaataaaaa
     4081 gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa aatctacaca
     4141 atgttctgtg tacacttctt atgtttttta cttctgataa atttttttga aacatcatag
     4201 aaaaaaccgc acacaaaata ccttatcata tgttacgttt cagtttatga ccgcaatttt
     4261 tatttcttcg cacgtctggg cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt
     4321 tggagtattt ttggaatttt tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt
     4381 tgctttttgg ggtttcccct attgtttgtc aagatttcga ggacggcgtt tttcttgcta
     4441 aaatcacaag tattgatgag cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg
     4501 ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg agtagtgtct atggggtttt
     4561 tgccttaaat gacagaatac attcccaata taccaaacat aactgtttcc tactagtcgg
     4621 ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4681 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4741 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4801 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4861 aaataccgca tcaggcggcc ttaagggcct cgtgatacgc ctatttttat aggttaatgt
     4921 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4981 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5041 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5101 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5161 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5221 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5281 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5341 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5401 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5461 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5521 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5581 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5641 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5701 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5761 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5821 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5881 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5941 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6001 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6061 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6121 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6181 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6241 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6301 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6361 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6421 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6481 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6541 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6601 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6661 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6721 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6781 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6841 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6901 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6961 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     7021 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     7081 cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat gatcttacta
     7141 actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac tcacaacgat
     7201 ggatacgcta acaacttgga aatgaaat
//
