LOCUS       sequence-87-expo        6787 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 6787)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6787)
  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: pPD93_97
FEATURES             Location/Qualifiers
     source          1..6787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2535..2555
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2598..2626)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3374..3394
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4361..4383)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4483..4502
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4580..4598)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4666..4770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4771..4839,4840..5631)
                     /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(4989..5008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5802..6390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6291..6310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6544..6561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6678..6699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6714..6719,6720..6737,6738..6744)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6752..6768
                     /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     6757..6779
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6776..6787,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6776..6787,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgggctgc aggtcggcta taataagttc ttgaataaaa
       61 taattttccc gacaaaacat gagtatttct ttcgaaaata aaagtgcagg ctaattagag
      121 attattctgt aattaactgc ataatttgtc acgtgccata gttttacatt ccactacgtc
      181 atagttctta aaatactaat ctcctgaaaa tagaagtagg tgaagaaagt ttaattatca
      241 gttctaaaat gacaattgat ctttggaata tgttctgaaa ctaccgatca ttgaacagat
      301 gctatttgaa tgatatagaa ttgtatattt gcaatttctg aaacgcgttc ttaaaggcac
      361 acagattaat tcaaaagggt ctggccgcaa aaaggtttat ggtggccgat tttgagtttt
      421 gtgtgtgatt gctttttcac aatcagtgtt ttcaggatta tgtgatgaac tagatcttca
      481 agtttcgtta catttcatat gttttcggaa ctcacgaagt acatattggg tattgtgctc
      541 aaaaaattca gcaatcagct tcgctccgct gactttagaa cccaaaaaaa tagtatggcc
      601 aaactgactg tgttacgatc atttcaattt ttcaatacat atttaagatt tctaagagta
      661 agaaggtcaa aaactgttct ggaatacata tatatttttc aggttacaat tagtcaaaaa
      721 gtgcactgaa atatacgttt taatttcacg aataacccaa ttagttcaat gtatttttgg
      781 tcaaccaacg ttaaagtttg gcttccaacc aattatcatt tctgatcaac cacaatgttt
      841 tttctttatc tgcaagttaa ttttttattt ttatccagat gtttggcata tttttcaatt
      901 cttcactagc gcccacttct tgcacttccg gcgccctgaa tctaatgcat ctgttgcaag
      961 aattgaaaga ccaatcaaca cattgttttc ttcacgagat actgaagaaa atgaataaaa
     1021 acagagaaaa agagccatgt gattagtgac aactgttgct aacagatacc atagcttgga
     1081 cttggtacgt gatggcaacg tatgggtcaa caaaaatgat tgcagagggg gtgcaaaaca
     1141 gtcaagtcga gaaaatatga aaaacagaaa acaaagaaca gaaaaatggg tttgagagtc
     1201 agtataattt ataaaagaaa aattgtacat agaaattaac catttttgta gaagaagtta
     1261 tttttcaagc atcgttaaaa attattcaaa gcaccttatt tcatatttaa ttttaaacat
     1321 ggttaaatga acaacacggt gcgcaatcag gaaaacttga aatctgaaac tgttgttgtg
     1381 atcttcttcg caactgttca gatagcacta gtgtaatgtt aagagtgcgc gaatataatg
     1441 gaatataatg gatcacacct cctgccatca ggtaaacgtc tctgttatca catatttcca
     1501 actattaaat ttttaccttt tacagtttta catttttttg aaaaaagtaa ctttttgtct
     1561 tcaaaatccc tgacgaaaat atcaaatatt ttaatcgaga ctgcagagga accgattgat
     1621 gatttggaaa atccagcttt acctgtgtaa gaactgaaaa gtttcataac cctagggtat
     1681 tcccagttac attccccact ggctaacaat agcacccagt ttttcatcac ctttcttcaa
     1741 atttctcggc gatttgttaa aaacaaaatt tgtgtccctt ctctgatatc tctatgtctc
     1801 taaacacaag ttcatcggaa aacgaaggag ggtaggtgtt ggttgggctc ccgaagtgaa
     1861 aatagaagag caagaataga atattagaga gagagtgcag agagggcggg atagctcccg
     1921 ggattccgtt ttcttcttct ttatcttcaa cgatgatgtg tgtgcgtgtt gtatagattc
     1981 tgttgctccc ccacaactcg ctccgaaggc tcaatacaat tcaattgata ttggaggaga
     2041 gcctaccgga gtgggaggat aagaagaaac ataagaagaa gaagaagaag aagcatgctt
     2101 ctggtttttg atgctatgaa aacggcacaa aaagatgatt gaggtccctt ttcaatacct
     2161 tctctcatct ttcaaatccc attgaaacct aaaacttctc accacgcttt accattgttc
     2221 tccaaaaact tatagcaatg tctataactt ttttatctct gaaaagcagt gttccatttt
     2281 tctttttcct attttatttc aattgtttct cacatttcgt ttggattctt tgcttgtcaa
     2341 ccagcttctt cttccacttt taccgtctaa ttttcagggc agggagccat caaacccacg
     2401 accactagat ccatctagag gatccccggg attggccaaa ggacccaaag gtatgtttcg
     2461 aatgatacta acataacata gaacattttc aggaggaccc ttggagggta ccgagctcag
     2521 aaaaaatgac tgctccaaag aagaagcgta aggtaccggt agaaaaaatg agtaaaggag
     2581 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     2641 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     2701 aatttatttg cactactgga aaactacctg ttccatgggt aagtttaaac atatatatac
     2761 taactaaccc tgattattta aattttcagc caacacttgt cactactttc tgttatggtg
     2821 ttcaatgctt ctcgagatac ccagatcata tgaaacggca tgactttttc aagagtgcca
     2881 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     2941 cacgtaagtt taaacagttc ggtactaact aaccatacat atttaaattt tcaggtgctg
     3001 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     3061 aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat
     3121 acatcatggc agacaaacaa aagaatggaa tcaaagttgt aagtttaaac atgattttac
     3181 taactaacta atctgattta aattttcaga acttcaaaat tagacacaac attgaagatg
     3241 gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
     3301 ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa
     3361 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg
     3421 atgaactata caaatagcat tcgtagaatt ccaactgagc gccggtcgct accattacca
     3481 acttgtctgg tgtcaaaaat aataggggcc gctgtcatca gagtaagttt aaactgagtt
     3541 ctactaacta acgagtaata tttaaatttt cagcatctcg cgcccgtgcc tctgacttct
     3601 aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat
     3661 ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac
     3721 ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa
     3781 aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac acaatgttct
     3841 gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac
     3901 cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct
     3961 tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta
     4021 tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt
     4081 tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac
     4141 aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg
     4201 gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta
     4261 aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg
     4321 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4381 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4441 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4501 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4561 gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4621 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4681 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4741 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4801 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4861 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4921 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4981 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5041 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5101 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5161 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5221 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5281 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5341 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5401 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5461 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5521 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5581 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5641 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5701 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5761 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5821 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5881 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5941 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6001 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6061 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6121 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6181 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6241 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6301 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6361 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6421 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6481 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6541 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6601 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6661 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6721 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6781 aacagct
//
