LOCUS       sequence-116-exp        8115 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 8115)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 8115)
  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: pPD96_62
FEATURES             Location/Qualifiers
     source          1..8115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             150..170
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(213..241)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     989..1009
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1142..1164)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5555..5577)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5677..5696
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5774..5792)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5860..5964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5965..6033,6034..6825)
                     /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(6183..6202)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6996..7584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7485..7504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7738..7755
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7872..7893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7908..7913,7914..7931,7932..7938)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7946..7962
                     /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     7951..7973
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7970..7986
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7970..7986
                     /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 cttgcttgga
      121 gggtaccgag ctcagaaaaa atgactgctc caaagaagaa gcgtaaggta ccggtagaaa
      181 aaatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
      241 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
      301 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tgggtaagtt
      361 taaacatata tatactaact aaccctgatt atttaaattt tcagccaaca cttgtcacta
      421 ctttctgtta tggtgttcaa tgcttctcga gatacccaga tcatatgaaa cggcatgact
      481 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
      541 acgggaacta caagacacgt aagtttaaac agttcggtac taactaacca tacatattta
      601 aattttcagg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
      661 aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata
      721 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt
      781 taaacatgat tttactaact aactaatctg atttaaattt tcagaacttc aaaattagac
      841 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
      901 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
      961 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     1021 ttacacatgg catggatgaa ctatacaaat gcccggtggg tgaagaccag aaacagcacc
     1081 tcgaactgag ccgcgatatt gcccagcgtt tcaacgcgct gtatggcgag atcgatcccg
     1141 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     1201 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1261 aacagttgcg aaggtaagtt taaacagatc catactaact aacttgttct gacataattt
     1321 tcagcttgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
     1381 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
     1441 tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc
     1501 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
     1561 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttaactcg gcgtttcatc
     1621 tgtggtgcaa cgggcgctgg gtcggttacg gccaggtaag tttaattaag ttgatactaa
     1681 ctaacaagat ctgattaatt ttcaggacag tcgtttgccg tctgaatttg acctgagcgc
     1741 atttttacgc gccggagaaa accgcctcgc ggtgatggtg ctgcgctgga gtgacggcag
     1801 ttatctggaa gatcaggata tgtggcggat gagcggcatt ttccgtgacg tctcgttgct
     1861 gcataaaccg actacacaaa tcagcgattt ccatgttgcc actcgcttta atgatgattt
     1921 ctcccgcgct gtactggagg ctgaagtcca ggtaagttta aacaggatct tactaactaa
     1981 catgctaaca ctgaattttc agatgtgcgg cgagttgcgt gactacctac gggtaacagt
     2041 ttctttatgg cagggtgaaa cgcaggtcgc cagcggcacc gcgcctttcg gcggtgaaat
     2101 tatcgatgag cgtggtggtt atgccgatcg cgtcacacta cgtctgaacg tcgaaaaccc
     2161 gaaactgtgg agcgccgaaa tcccgaatct ctatcgtgcg gtggttgaac tgcacaccgc
     2221 cgacggcacg ctgattgaag cagaagcctg cgatgtcggt ttccgcgagg tgcggattga
     2281 aaatggtctg ctgctgctga acggcaagcc gttgctgatt cgaggcgtta accgtcacga
     2341 gcatcatcct ctgcatggtc aggtcatgga tgagcagacg atggtgcagg atgtaagttt
     2401 aaactattcg ttactaacta actttaaaca tttaaatttt cagatcctgc tgatgaagca
     2461 gaacaacttt aacgccgtgc gctgttcgca ttatccgaac catccgctgt ggtacacgct
     2521 gtgcgaccgc tacggcctgt atgtggtgga tgaagccaat attgaaaccc acggcatggt
     2581 gccaatgaat cgtctgaccg atgatccgcg ctggctaccg gcgatgagcg aacgcgtaac
     2641 gcgaatggtg cagcgcgatc gtaatcaccc gagtgtgatc atctggtcgc tggggaatgg
     2701 taagtttaaa cagttgaata ctaactaacg gagatctttg aaattttcag aatcaggcca
     2761 cggcgctaat cacgacgcgc tgtatcgctg gatcaaatct gtcgatcctt cccgcccggt
     2821 gcagtatgaa ggcggcggag ccgacaccac ggccaccgat attatttgcc cgatgtacgc
     2881 gcgcgtggat gaagaccagc ccttcccggc tgtgccgaaa tggtccatca aaaaatggct
     2941 ttcgctacct ggagagacgc gcccgctgat tctttgcgag gtaagtttaa acagaactct
     3001 actaactaac acattagatc ctaattttca gtacgctcac gcgatgggca acagtcttgg
     3061 cggtttcgct aaatactggc aggcgtttcg tcagtatccc cgtttacagg gcggcttcgt
     3121 ctgggactgg gtggatcagt cgctgattaa atatgatgaa aacggcaacc cgtggtcggc
     3181 ttacggcggt gattttggcg atacgccgaa cgatcgccag ttctgtatga acggtctggt
     3241 ctttgccgac cgcacgccgc atccagcgta agtttaaaca ataacctact aactaacgta
     3301 gataatttaa attttcaggc tgacggaagc aaaacaccag cagcagtttt tccagttccg
     3361 tttatccggg caaaccatcg aagtgaccag cgaatacctg ttccgtcata gcgataacga
     3421 gctcctgcac tggatggtgg cgctggatgg taagccgctg gcaagcggtg aagtgcctct
     3481 ggatgtcgct ccacaaggta aacagttgat tgaactgcct gaactaccgc agccggagag
     3541 cgccgggcaa ctctggctca cagtacgcgt agtgcaaccg aacgcgaccg catggtcaga
     3601 agccgggcac atcagcgcat ggcagcagtg gaggtaagtt taaacaagat cctactaact
     3661 aactctacat tgatgaattt tcagactggc ggaaaacctc agtgtgacgc tccccgccgc
     3721 gtcccacgcc atcccgcatc tgaccaccag cgaaatggat ttttgcatcg agctgggtaa
     3781 taagcgttgg caatttaacc gccagtcagg ctttctttca cagatgtgga ttggcgataa
     3841 aaaacaactg ctgacgccgc tgcgcgatca gttcacccgt gcaccgctgg ataacgacat
     3901 tggcgtaagt gaagcgaccc gcattgaccc taacgcctgg gtcggtaagt ttaaacaaag
     3961 ttgtactaac taacgaagat cttgataatt ttcagaacgc tggaaggcgg cgggccatta
     4021 ccaggccgaa gcagcgttgt tgcagtgcac ggcagataca cttgctgatg cggtgctgat
     4081 tacgaccgct cacgcgtggc agcatcaggg gaaaacctta tttatcagcc ggaaaaccta
     4141 ccggattgat ggtagtggtc aaatggcgat taccgttgat gttgaagtgg cgagcgatac
     4201 accgcatccg gcgcggattg gcctgaactg ccagctggcg caggtagcag agcgggtaaa
     4261 ctggctcgga ttagggccgc aagaaaacta tcccgaccgc cttactgccg cctgttttga
     4321 ccgctgggat ctgccattgt cagacatgta gtaagtttaa acttgatagt actaactaac
     4381 atgtttcatt taaattttca gtaccccgta cgtcttcccg agcgaaaacg gtctgcgctg
     4441 cgggacgcgc gaattgaatt atggcccaca ccagtggcgc ggcgacttcc agttcaacat
     4501 cagccgctac agtcaacagc aactgatgga aaccagccat cgccatctgc tgcacgcgga
     4561 agaaggcaca tggctgaata tcgacggttt ccatatgggg attggtggcg acgactcctg
     4621 gagcccgtca gtatcggcgg aattccaact gagcgccggt cgctaccatt accaacttgt
     4681 ctggtgtcaa aaataatagg ggccgctgtc atcagagtaa gtttaaactg agttctacta
     4741 actaacgagt aatatttaaa ttttcagcat ctcgcgcccg tgcctctgac ttctaagtcc
     4801 aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc ctatttttgt
     4861 tattatcaaa aaacttctct taatttcttt gttttttagc ttcttttaag tcacctctaa
     4921 caatgaaatt gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt
     4981 gtgctccctc cccccattaa taataattct atcccaaaat ctacacaatg ttctgtgtac
     5041 acttcttatg ttttttactt ctgataaatt tttttgaaac atcatagaaa aaaccgcaca
     5101 caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat ttcttcgcac
     5161 gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg agtatttttg
     5221 gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc tttttggggt
     5281 ttcccctatt gtttgtcaag atttcgagga cggcgttttt cttgctaaaa tcacaagtat
     5341 tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaaggt
     5401 gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc cttaaatgac
     5461 agaatacatt cccaatatac caaacataac tgtttcctac tagtcggccg tacgggccct
     5521 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5581 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5641 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     5701 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     5761 ggcggcctta agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5821 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5881 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5941 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6001 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6061 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6121 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6181 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6241 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6301 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6361 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6421 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6481 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6541 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6601 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6661 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6721 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6781 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6841 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6901 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6961 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7021 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7081 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7141 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7201 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7261 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7321 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7381 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7441 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7501 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7561 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7621 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7681 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7741 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7801 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7861 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7921 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7981 tatgaccatg attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc
     8041 atttaaattt tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca
     8101 acttggaaat gaaat
//
