LOCUS       sequence-111-exp        7661 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 7661)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7661)
  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_93
FEATURES             Location/Qualifiers
     source          1..7661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             386..406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(449..477)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1225..1245
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1376..4423
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(1378..1400)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5235..5257)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5357..5376
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5454..5472)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5540..5644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5645..5713,5714..6505)
                     /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(5863..5882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6676..7264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7165..7184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7418..7435
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7552..7573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7588..7593,7594..7611,7612..7618)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7626..7642
                     /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     7631..7653
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7650..7661,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7650..7661,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccaattc
       61 ttgagacttc cactatcttc tctggcaccc tccactacgc gtaggcgtct ctcgcttcgt
      121 gtattcccgg gaagccgggt tcccgtctct cccgccgctg ccgctgccgc acacagcttt
      181 acacctcgta gaatccccaa agaggggcgt ggcttgcggg tgccaacatc ctcctgccga
      241 ggaagaagca ggcactcatc actcgcatca tcaacctcgg gattggccaa aggacccaaa
      301 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      361 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tagaaaaaat
      421 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
      481 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
      541 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa
      601 catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt
      661 ctgttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc atgacttttt
      721 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
      781 gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
      841 ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
      901 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
      961 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa
     1021 catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa
     1081 cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga
     1141 tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga
     1201 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     1261 acatggcatg gatgaactat acaaatgccc ggtgggtgaa gaccagaaac agcacctcga
     1321 actgagccgc gatattgccc agcgtttcaa cgcgctgtat ggcgagatcg atcccgtcgt
     1381 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1441 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1501 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
     1561 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
     1621 gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa
     1681 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     1741 tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgtta actcggcgtt
     1801 tcatctgtgg tgcaacgggc gctgggtcgg ttacggccag gacagtcgtt tgccgtctga
     1861 atttgacctg agcgcatttt tacgcgccgg agaaaaccgc ctcgcggtga tggtgctgcg
     1921 ctggagtgac ggcagttatc tggaagatca ggatatgtgg cggatgagcg gcattttccg
     1981 tgacgtctcg ttgctgcata aaccgactac acaaatcagc gatttccatg ttgccactcg
     2041 ctttaatgat gatttcagcc gcgctgtact ggaggctgaa gttcagatgt gcggcgagtt
     2101 gcgtgactac ctacgggtaa cagtttcttt atggcagggt gaaacgcagg tcgccagcgg
     2161 caccgcgcct ttcggcggtg aaattatcga tgagcgtggt ggttatgccg atcgcgtcac
     2221 actacgtctg aacgtcgaaa acccgaaact gtggagcgcc gaaatcccga atctctatcg
     2281 tgcggtggtt gaactgcaca ccgccgacgg cacgctgatt gaagcagaag cctgcgatgt
     2341 cggtttccgc gaggtgcgga ttgaaaatgg tctgctgctg ctgaacggca agccgttgct
     2401 gattcgaggc gttaaccgtc acgagcatca tcctctgcat ggtcaggtca tggatgagca
     2461 gacgatggtg caggatatcc tgctgatgaa gcagaacaac tttaacgccg tgcgctgttc
     2521 gcattatccg aaccatccgc tgtggtacac gctgtgcgac cgctacggcc tgtatgtggt
     2581 ggatgaagcc aatattgaaa cccacggcat ggtgccaatg aatcgtctga ccgatgatcc
     2641 gcgctggcta ccggcgatga gcgaacgcgt aacgcgaatg gtgcagcgcg atcgtaatca
     2701 cccgagtgtg atcatctggt cgctggggaa tgaatcaggc cacggcgcta atcacgacgc
     2761 gctgtatcgc tggatcaaat ctgtcgatcc ttcccgcccg gtgcagtatg aaggcggcgg
     2821 agccgacacc acggccaccg atattatttg cccgatgtac gcgcgcgtgg atgaagacca
     2881 gcccttcccg gctgtgccga aatggtccat caaaaaatgg ctttcgctac ctggagagac
     2941 gcgcccgctg atcctttgcg aatacgccca cgcgatgggt aacagtcttg gcggtttcgc
     3001 taaatactgg caggcgtttc gtcagtatcc ccgtttacag ggcggcttcg tctgggactg
     3061 ggtggatcag tcgctgatta aatatgatga aaacggcaac ccgtggtcgg cttacggcgg
     3121 tgattttggc gatacgccga acgatcgcca gttctgtatg aacggtctgg tctttgccga
     3181 ccgcacgccg catccagcgc tgacggaagc aaaacaccag cagcagtttt tccagttccg
     3241 tttatccggg caaaccatcg aagtgaccag cgaatacctg ttccgtcata gcgataacga
     3301 gctcctgcac tggatggtgg cgctggatgg taagccgctg gcaagcggtg aagtgcctct
     3361 ggatgtcgct ccacaaggta aacagttgat tgaactgcct gaactaccgc agccggagag
     3421 cgccgggcaa ctctggctca cagtacgcgt agtgcaaccg aacgcgaccg catggtcaga
     3481 agccgggcac atcagcgcct ggcagcagtg gcgtctggcg gaaaacctca gtgtgacgct
     3541 ccccgccgcg tcccacgcca tcccgcatct gaccaccagc gaaatggatt tttgcatcga
     3601 gctgggtaat aagcgttggc aatttaaccg ccagtcaggc tttctttcac agatgtggat
     3661 tggcgataaa aaacaactgc tgacgccgct gcgcgatcag ttcacccgtg caccgctgga
     3721 taacgacatt ggcgtaagtg aagcgacccg cattgaccct aacgcctggg tcgaacgctg
     3781 gaaggcggcg ggccattacc aggccgaagc agcgttgttg cagtgcacgg cagatacact
     3841 tgctgatgcg gtgctgatta cgaccgctca cgcgtggcag catcagggga aaaccttatt
     3901 tatcagccgg aaaacctacc ggattgatgg tagtggtcaa atggcgatta ccgttgatgt
     3961 tgaagtggcg agcgatacac cgcatccggc gcggattggc ctgaactgcc agctggcgca
     4021 ggtagcagag cgggtaaact ggctcggatt agggccgcaa gaaaactatc ccgaccgcct
     4081 tactgccgcc tgttttgacc gctgggatct gccattgtca gacatgtata ccccgtacgt
     4141 cttcccgagc gaaaacggtc tgcgctgcgg gacgcgcgaa ttgaattatg gcccacacca
     4201 gtggcgcggc gacttccagt tcaacatcag ccgctacagt caacagcaac tgatggaaac
     4261 cagccatcgc catctgctgc acgcggaaga aggcacatgg ctgaatatcg acggtttcca
     4321 tatggggatt ggtggcgacg actcctggag cccgtcagta tcggcggaat tccaactgag
     4381 cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc
     4441 agatcgccat ctcgcgcccg tgcctctgac ttctaagtcc aattactctt caacatccct
     4501 acatgctctt tctccctgtg ctcccacccc ctatttttgt tattatcaaa aaacttctct
     4561 taatttcttt gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc
     4621 aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa
     4681 taataattct atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttactt
     4741 ctgataaatt tttttgaaac atcatagaaa aaaccgcaca caaaatacct tatcatatgt
     4801 tacgtttcag tttatgaccg caatttttat ttcttcgcac gtctgggcct ctcatgacgt
     4861 caaatcatgc tcatcgtgaa aaagttttgg agtatttttg gaatttttca atcaagtgaa
     4921 agtttatgaa attaattttc ctgcttttgc tttttggggt ttcccctatt gtttgtcaag
     4981 atttcgagga cggcgttttt cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa
     5041 agatcggaag aaggtttggg tttgaggctc agtggaaggt gagtagaagt tgataatttg
     5101 aaagtggagt agtgtctatg gggtttttgc cttaaatgac agaatacatt cccaatatac
     5161 caaacataac tgtttcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt
     5221 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5281 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5341 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5401 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt
     5461 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5521 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5581 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5641 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5701 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5761 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5821 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5881 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5941 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6001 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6061 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6121 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6181 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6241 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6301 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6361 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6421 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6481 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6541 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6601 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6661 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6721 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6781 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6841 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6901 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6961 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7021 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc
     7081 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7141 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7201 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7261 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7321 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7381 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7441 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     7501 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     7561 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     7621 tggaattgtg agcggataac aatttcacac aggaaacagc t
//
