LOCUS       sequence-113-exp        8111 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 8111)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 8111)
  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_04
FEATURES             Location/Qualifiers
     source          1..8111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             146..166
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(209..237)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     985..1005
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1138..1160)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5551..5573)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5673..5692
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5770..5788)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5856..5960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5961..6029,6030..6821)
                     /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(6179..6198)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6992..7580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7481..7500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7734..7751
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7868..7889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7904..7909,7910..7927,7928..7934)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7942..7958
                     /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     7947..7969
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7966..7982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7966..7982
                     /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 cttggagggt
      121 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tagaaaaaat
      181 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
      241 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
      301 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa
      361 catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt
      421 ctgttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc atgacttttt
      481 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
      541 gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
      601 ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
      661 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
      721 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa
      781 catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa
      841 cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga
      901 tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga
      961 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     1021 acatggcatg gatgaactat acaaatgccc ggtgggtgaa gaccagaaac agcacctcga
     1081 actgagccgc gatattgccc agcgtttcaa cgcgctgtat ggcgagatcg atcccgtcgt
     1141 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1201 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1261 gttgcgaagg taagtttaaa cagatccata ctaactaact tgttctgaca taattttcag
     1321 cttgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg
     1381 gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca
     1441 cggttacgat gcgcccatct acaccaacgt gacctatccc attacggtca atccgccgtt
     1501 tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg
     1561 gctacaggaa ggccagacgc gaattatttt tgatggcgtt aactcggcgt ttcatctgtg
     1621 gtgcaacggg cgctgggtcg gttacggcca ggtaagttta attaagttga tactaactaa
     1681 caagatctga ttaattttca ggacagtcgt ttgccgtctg aatttgacct gagcgcattt
     1741 ttacgcgccg gagaaaaccg cctcgcggtg atggtgctgc gctggagtga cggcagttat
     1801 ctggaagatc aggatatgtg gcggatgagc ggcattttcc gtgacgtctc gttgctgcat
     1861 aaaccgacta cacaaatcag cgatttccat gttgccactc gctttaatga tgatttctcc
     1921 cgcgctgtac tggaggctga agtccaggta agtttaaaca ggatcttact aactaacatg
     1981 ctaacactga attttcagat gtgcggcgag ttgcgtgact acctacgggt aacagtttct
     2041 ttatggcagg gtgaaacgca ggtcgccagc ggcaccgcgc ctttcggcgg tgaaattatc
     2101 gatgagcgtg gtggttatgc cgatcgcgtc acactacgtc tgaacgtcga aaacccgaaa
     2161 ctgtggagcg ccgaaatccc gaatctctat cgtgcggtgg ttgaactgca caccgccgac
     2221 ggcacgctga ttgaagcaga agcctgcgat gtcggtttcc gcgaggtgcg gattgaaaat
     2281 ggtctgctgc tgctgaacgg caagccgttg ctgattcgag gcgttaaccg tcacgagcat
     2341 catcctctgc atggtcaggt catggatgag cagacgatgg tgcaggatgt aagtttaaac
     2401 tattcgttac taactaactt taaacattta aattttcaga tcctgctgat gaagcagaac
     2461 aactttaacg ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta cacgctgtgc
     2521 gaccgctacg gcctgtatgt ggtggatgaa gccaatattg aaacccacgg catggtgcca
     2581 atgaatcgtc tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg cgtaacgcga
     2641 atggtgcagc gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg gaatggtaag
     2701 tttaaacagt tgaatactaa ctaacggaga tctttgaaat tttcagaatc aggccacggc
     2761 gctaatcacg acgcgctgta tcgctggatc aaatctgtcg atccttcccg cccggtgcag
     2821 tatgaaggcg gcggagccga caccacggcc accgatatta tttgcccgat gtacgcgcgc
     2881 gtggatgaag accagccctt cccggctgtg ccgaaatggt ccatcaaaaa atggctttcg
     2941 ctacctggag agacgcgccc gctgattctt tgcgaggtaa gtttaaacag aactctacta
     3001 actaacacat tagatcctaa ttttcagtac gctcacgcga tgggcaacag tcttggcggt
     3061 ttcgctaaat actggcaggc gtttcgtcag tatccccgtt tacagggcgg cttcgtctgg
     3121 gactgggtgg atcagtcgct gattaaatat gatgaaaacg gcaacccgtg gtcggcttac
     3181 ggcggtgatt ttggcgatac gccgaacgat cgccagttct gtatgaacgg tctggtcttt
     3241 gccgaccgca cgccgcatcc agcgtaagtt taaacaataa cctactaact aacgtagata
     3301 atttaaattt tcaggctgac ggaagcaaaa caccagcagc agtttttcca gttccgttta
     3361 tccgggcaaa ccatcgaagt gaccagcgaa tacctgttcc gtcatagcga taacgagctc
     3421 ctgcactgga tggtggcgct ggatggtaag ccgctggcaa gcggtgaagt gcctctggat
     3481 gtcgctccac aaggtaaaca gttgattgaa ctgcctgaac taccgcagcc ggagagcgcc
     3541 gggcaactct ggctcacagt acgcgtagtg caaccgaacg cgaccgcatg gtcagaagcc
     3601 gggcacatca gcgcatggca gcagtggagg taagtttaaa caagatccta ctaactaact
     3661 ctacattgat gaattttcag actggcggaa aacctcagtg tgacgctccc cgccgcgtcc
     3721 cacgccatcc cgcatctgac caccagcgaa atggattttt gcatcgagct gggtaataag
     3781 cgttggcaat ttaaccgcca gtcaggcttt ctttcacaga tgtggattgg cgataaaaaa
     3841 caactgctga cgccgctgcg cgatcagttc acccgtgcac cgctggataa cgacattggc
     3901 gtaagtgaag cgacccgcat tgaccctaac gcctgggtcg gtaagtttaa acaaagttgt
     3961 actaactaac gaagatcttg ataattttca gaacgctgga aggcggcggg ccattaccag
     4021 gccgaagcag cgttgttgca gtgcacggca gatacacttg ctgatgcggt gctgattacg
     4081 accgctcacg cgtggcagca tcaggggaaa accttattta tcagccggaa aacctaccgg
     4141 attgatggta gtggtcaaat ggcgattacc gttgatgttg aagtggcgag cgatacaccg
     4201 catccggcgc ggattggcct gaactgccag ctggcgcagg tagcagagcg ggtaaactgg
     4261 ctcggattag ggccgcaaga aaactatccc gaccgcctta ctgccgcctg ttttgaccgc
     4321 tgggatctgc cattgtcaga catgtagtaa gtttaaactt gatagtacta actaacatgt
     4381 ttcatttaaa ttttcagtac cccgtacgtc ttcccgagcg aaaacggtct gcgctgcggg
     4441 acgcgcgaat tgaattatgg cccacaccag tggcgcggcg acttccagtt caacatcagc
     4501 cgctacagtc aacagcaact gatggaaacc agccatcgcc atctgctgca cgcggaagaa
     4561 ggcacatggc tgaatatcga cggtttccat atggggattg gtggcgacga ctcctggagc
     4621 ccgtcagtat cggcggaatt ccaactgagc gccggtcgct accattacca acttgtctgg
     4681 tgtcaaaaat aataggggcc gctgtcatca gagtaagttt aaactgagtt ctactaacta
     4741 acgagtaata tttaaatttt cagcatctcg cgcccgtgcc tctgacttct aagtccaatt
     4801 actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt
     4861 atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat
     4921 gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc
     4981 tccctccccc cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt
     5041 cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa
     5101 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct
     5161 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat
     5221 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc
     5281 cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     5341 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     5401 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     5461 tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg
     5521 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5581 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5641 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5701 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5761 gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5821 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5881 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5941 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6001 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6061 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6121 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6181 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6241 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6301 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6361 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6421 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6481 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6541 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6601 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6661 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6721 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6781 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6841 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6901 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6961 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7021 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7081 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7141 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7201 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7261 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7321 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7381 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7441 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7501 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7561 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7621 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7681 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7741 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     7801 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     7861 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     7921 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     7981 accatgatta cgccaagctg taagtttaaa catgatctta ctaactaact attctcattt
     8041 aaattttcag agcttaaaaa tggctgaaat cactcacaac gatggatacg ctaacaactt
     8101 ggaaatgaaa t
//
