LOCUS       sequence-100-exp        7390 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 7390)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7390)
  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_62
FEATURES             Location/Qualifiers
     source          1..7390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             307..327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(417..439)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4830..4852)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4952..4971
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5049..5067)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5135..5239
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5240..5308,5309..6100)
                     /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(5458..5477)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6271..6859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6760..6779
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7013..7030
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7147..7168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7183..7188,7189..7206,7207..7213)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7221..7237
                     /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     7226..7248
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7245..7261
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7245..7261
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagacact tttcagctac ctagatacat
       61 ggatatcccc gcctcccaat ccacccaccc agggaaaaag aagggctcgc cgaaaatcaa
      121 agttatctcc aggctcgcgc atcccaccga gcggttgact tctctccacc acttttcatt
      181 ttaaccctcg gggtacggga ttggccaaag gacccaaagg tatgtttcga atgatactaa
      241 cataacatag aacattttca ggaggaccct tcttggaggg taccgagctc agaaaaaatg
      301 actgctccaa agaagaagcg taaggtaccg gtgggtgaag accagaaaca gcacctcgaa
      361 ctgagccgcg atattgccca gcgtttcaac gcgctgtatg gcgagatcga tcccgtcgtt
      421 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
      481 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
      541 ttgcgaaggt aagtttaaac agatccatac taactaactt gttctgacat aattttcagc
      601 ttgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg
      661 ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac
      721 ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt
      781 gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg
      841 ctacaggaag gccagacgcg aattattttt gatggcgtta actcggcgtt tcatctgtgg
      901 tgcaacgggc gctgggtcgg ttacggccag gtaagtttaa ttaagttgat actaactaac
      961 aagatctgat taattttcag gacagtcgtt tgccgtctga atttgacctg agcgcatttt
     1021 tacgcgccgg agaaaaccgc ctcgcggtga tggtgctgcg ctggagtgac ggcagttatc
     1081 tggaagatca ggatatgtgg cggatgagcg gcattttccg tgacgtctcg ttgctgcata
     1141 aaccgactac acaaatcagc gatttccatg ttgccactcg ctttaatgat gatttctccc
     1201 gcgctgtact ggaggctgaa gtccaggtaa gtttaaacag gatcttacta actaacatgc
     1261 taacactgaa ttttcagatg tgcggcgagt tgcgtgacta cctacgggta acagtttctt
     1321 tatggcaggg tgaaacgcag gtcgccagcg gcaccgcgcc tttcggcggt gaaattatcg
     1381 atgagcgtgg tggttatgcc gatcgcgtca cactacgtct gaacgtcgaa aacccgaaac
     1441 tgtggagcgc cgaaatcccg aatctctatc gtgcggtggt tgaactgcac accgccgacg
     1501 gcacgctgat tgaagcagaa gcctgcgatg tcggtttccg cgaggtgcgg attgaaaatg
     1561 gtctgctgct gctgaacggc aagccgttgc tgattcgagg cgttaaccgt cacgagcatc
     1621 atcctctgca tggtcaggtc atggatgagc agacgatggt gcaggatgta agtttaaact
     1681 attcgttact aactaacttt aaacatttaa attttcagat cctgctgatg aagcagaaca
     1741 actttaacgc cgtgcgctgt tcgcattatc cgaaccatcc gctgtggtac acgctgtgcg
     1801 accgctacgg cctgtatgtg gtggatgaag ccaatattga aacccacggc atggtgccaa
     1861 tgaatcgtct gaccgatgat ccgcgctggc taccggcgat gagcgaacgc gtaacgcgaa
     1921 tggtgcagcg cgatcgtaat cacccgagtg tgatcatctg gtcgctgggg aatggtaagt
     1981 ttaaacagtt gaatactaac taacggagat ctttgaaatt ttcagaatca ggccacggcg
     2041 ctaatcacga cgcgctgtat cgctggatca aatctgtcga tccttcccgc ccggtgcagt
     2101 atgaaggcgg cggagccgac accacggcca ccgatattat ttgcccgatg tacgcgcgcg
     2161 tggatgaaga ccagcccttc ccggctgtgc cgaaatggtc catcaaaaaa tggctttcgc
     2221 tacctggaga gacgcgcccg ctgattcttt gcgaggtaag tttaaacaga actctactaa
     2281 ctaacacatt agatcctaat tttcagtacg ctcacgcgat gggcaacagt cttggcggtt
     2341 tcgctaaata ctggcaggcg tttcgtcagt atccccgttt acagggcggc ttcgtctggg
     2401 actgggtgga tcagtcgctg attaaatatg atgaaaacgg caacccgtgg tcggcttacg
     2461 gcggtgattt tggcgatacg ccgaacgatc gccagttctg tatgaacggt ctggtctttg
     2521 ccgaccgcac gccgcatcca gcgtaagttt aaacaataac ctactaacta acgtagataa
     2581 tttaaatttt caggctgacg gaagcaaaac accagcagca gtttttccag ttccgtttat
     2641 ccgggcaaac catcgaagtg accagcgaat acctgttccg tcatagcgat aacgagctcc
     2701 tgcactggat ggtggcgctg gatggtaagc cgctggcaag cggtgaagtg cctctggatg
     2761 tcgctccaca aggtaaacag ttgattgaac tgcctgaact accgcagccg gagagcgccg
     2821 ggcaactctg gctcacagta cgcgtagtgc aaccgaacgc gaccgcatgg tcagaagccg
     2881 ggcacatcag cgcatggcag cagtggaggt aagtttaaac aagatcctac taactaactc
     2941 tacattgatg aattttcaga ctggcggaaa acctcagtgt gacgctcccc gccgcgtccc
     3001 acgccatccc gcatctgacc accagcgaaa tggatttttg catcgagctg ggtaataagc
     3061 gttggcaatt taaccgccag tcaggctttc tttcacagat gtggattggc gataaaaaac
     3121 aactgctgac gccgctgcgc gatcagttca cccgtgcacc gctggataac gacattggcg
     3181 taagtgaagc gacccgcatt gaccctaacg cctgggtcgg taagtttaaa caaagttgta
     3241 ctaactaacg aagatcttga taattttcag aacgctggaa ggcggcgggc cattaccagg
     3301 ccgaagcagc gttgttgcag tgcacggcag atacacttgc tgatgcggtg ctgattacga
     3361 ccgctcacgc gtggcagcat caggggaaaa ccttatttat cagccggaaa acctaccgga
     3421 ttgatggtag tggtcaaatg gcgattaccg ttgatgttga agtggcgagc gatacaccgc
     3481 atccggcgcg gattggcctg aactgccagc tggcgcaggt agcagagcgg gtaaactggc
     3541 tcggattagg gccgcaagaa aactatcccg accgccttac tgccgcctgt tttgaccgct
     3601 gggatctgcc attgtcagac atgtagtaag tttaaacttg atagtactaa ctaacatgtt
     3661 tcatttaaat tttcagtacc ccgtacgtct tcccgagcga aaacggtctg cgctgcggga
     3721 cgcgcgaatt gaattatggc ccacaccagt ggcgcggcga cttccagttc aacatcagcc
     3781 gctacagtca acagcaactg atggaaacca gccatcgcca tctgctgcac gcggaagaag
     3841 gcacatggct gaatatcgac ggtttccata tggggattgg tggcgacgac tcctggagcc
     3901 cgtcagtatc ggcggaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt
     3961 gtcaaaaata ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa
     4021 cgagtaatat ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta
     4081 ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta
     4141 tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg
     4201 aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct
     4261 ccctcccccc attaataata attctatccc aaaatctaca caatgttctg tgtacacttc
     4321 ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa
     4381 taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg
     4441 ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt
     4501 tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc
     4561 ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg
     4621 agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta
     4681 gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat
     4741 acattcccaa tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt
     4801 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     4861 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     4921 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     4981 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg
     5041 ccttaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5101 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5161 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5221 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5281 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5341 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5401 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5461 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5521 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5581 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5641 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5701 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5761 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5821 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5881 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5941 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6001 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6061 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6121 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6181 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6241 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6301 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6361 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     6421 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6481 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6541 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6601 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6661 gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6721 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6781 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6841 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6901 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6961 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     7021 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     7081 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     7141 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     7201 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     7261 ccatgattac gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta
     7321 aattttcaga gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg
     7381 gaaatgaaat
//
