LOCUS       sequence-72-expo        7131 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 7131)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7131)
  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: pPD88_27
FEATURES             Location/Qualifiers
     source          1..7131
                     /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(496..518)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4705..4727)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4827..4846
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4924..4942)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5010..5114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5115..5183,5184..5975)
                     /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(5333..5352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6146..6734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6635..6654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6888..6905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7022..7043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7058..7063,7064..7081,7082..7088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7096..7112
                     /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     7101..7123
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7120..7131,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7120..7131,1..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 tgggtgaaga
      421 ccagaaacag cacctcgaac tgagccgcga tattgcccag cgtttcaacg cgctgtatgg
      481 cgagatcgat cccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      541 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      601 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc
      661 ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt
      721 cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac
      781 ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc
      841 gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga
      901 tggcgttaac tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt acggccaggt
      961 aagtttaatt aagttgatac taactaacaa gatctgatta attttcagga cagtcgtttg
     1021 ccgtctgaat ttgacctgag cgcattttta cgcgccggag aaaaccgcct cgcggtgatg
     1081 gtgctgcgct ggagtgacgg cagttatctg gaagatcagg atatgtggcg gatgagcggc
     1141 attttccgtg acgtctcgtt gctgcataaa ccgactacac aaatcagcga tttccatgtt
     1201 gccactcgct ttaatgatga tttcagccgc gctgtactgg aggctgaagt tcagatgtgc
     1261 ggcgagttgc gtgactacct acgggtaaca gtttctttat ggcagggtga aacgcaggtc
     1321 gccagcggca ccgcgccttt cggcggtgaa attatcgatg agcgtggtgg ttatgccgat
     1381 cgcgtcacac tacgtctgaa cgtcgaaaac ccgaaactgt ggagcgccga aatcccgaat
     1441 ctctatcgtg cggtggttga actgcacacc gccgacggca cgctgattga agcagaagcc
     1501 tgcgatgtcg gtttccgcga ggtgcggatt gaaaatggtc tgctgctgct gaacggcaag
     1561 ccgttgctga ttcgaggcgt taaccgtcac gagcatcatc ctctgcatgg tcaggtcatg
     1621 gatgagcaga cgatggtgca ggatgtaagt ttaaactatt cgttactaac taactttaaa
     1681 catttaaatt ttcagatcct gctgatgaag cagaacaact ttaacgccgt gcgctgttcg
     1741 cattatccga accatccgct gtggtacacg ctgtgcgacc gctacggcct gtatgtggtg
     1801 gatgaagcca atattgaaac ccacggcatg gtgccaatga atcgtctgac cgatgatccg
     1861 cgctggctac cggcgatgag cgaacgcgta acgcgaatgg tgcagcgcga tcgtaatcac
     1921 ccgagtgtga tcatctggtc gctggggaat ggtaagttta aacagttgaa tactaactaa
     1981 cggagatctt tgaaattttc agaatcaggc cacggcgcta atcacgacgc gctgtatcgc
     2041 tggatcaaat ctgtcgatcc ttcccgcccg gtgcagtatg aaggcggcgg agccgacacc
     2101 acggccaccg atattatttg cccgatgtac gcgcgcgtgg atgaagacca gcccttcccg
     2161 gctgtgccga aatggtccat caaaaaatgg ctttcgctac ctggagagac gcgcccgctg
     2221 atcctttgcg aatacgccca cgcgatgggt aacagtcttg gcggtttcgc taaatactgg
     2281 caggcgtttc gtcagtatcc ccgtttacag ggcggcttcg tctgggactg ggtggatcag
     2341 tcgctgatta aatatgatga aaacggcaac ccgtggtcgg cttacggcgg tgattttggc
     2401 gatacgccga acgatcgcca gttctgtatg aacggtctgg tctttgccga ccgcacgccg
     2461 catccagcgt aagtttaaac aataacctac taactaacgt agataattta aattttcagg
     2521 ctgacggaag caaaacacca gcagcagttt ttccagttcc gtttatccgg gcaaaccatc
     2581 gaagtgacca gcgaatacct gttccgtcat agcgataacg agctcctgca ctggatggtg
     2641 gcgctggatg gtaagccgct ggcaagcggt gaagtgcctc tggatgtcgc tccacaaggt
     2701 aaacagttga ttgaactgcc tgaactaccg cagccggaga gcgccgggca actctggctc
     2761 acagtacgcg tagtgcaacc gaacgcgacc gcatggtcag aagccgggca catcagcgcc
     2821 tggcagcagt ggcgtctggc ggaaaacctc agtgtgacgc tccccgccgc gtcccacgcc
     2881 atcccgcatc tgaccaccag cgaaatggat ttttgcatcg agctgggtaa taagcgttgg
     2941 caatttaacc gccagtcagg ctttctttca cagatgtgga ttggcgataa aaaacaactg
     3001 ctgacgccgc tgcgcgatca gttcacccgt gcaccgctgg ataacgacat tggcgtaagt
     3061 gaagcgaccc gcattgaccc taacgcctgg gtcggtaagt ttaaacaaag ttgtactaac
     3121 taacgaagat cttgataatt ttcagaacgc tggaaggcgg cgggccatta ccaggccgaa
     3181 gcagcgttgt tgcagtgcac ggcagataca cttgctgatg cggtgctgat tacgaccgct
     3241 cacgcgtggc agcatcaggg gaaaacctta tttatcagcc ggaaaaccta ccggattgat
     3301 ggtagtggtc aaatggcgat taccgttgat gttgaagtgg cgagcgatac accgcatccg
     3361 gcgcggattg gcctgaactg ccagctggcg caggtagcag agcgggtaaa ctggctcgga
     3421 ttagggccgc aagaaaacta tcccgaccgc cttactgccg cctgttttga ccgctgggat
     3481 ctgccattgt cagacatgta gtaagtttaa acttgatagt actaactaac atgtttcatt
     3541 taaattttca gtaccccgta cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc
     3601 gaattgaatt atggcccaca ccagtggcgc ggcgacttcc agttcaacat cagccgctac
     3661 agtcaacagc aactgatgga aaccagccat cgccatctgc tgcacgcgga agaaggcaca
     3721 tggctgaata tcgacggttt ccatatgggg attggtggcg acgactcctg gagcccgtca
     3781 gtatcggcgg aattccaact gagcgccggt cgctaccatt accaacttgt ctggtgtcaa
     3841 aaataatagg ggccgctgtc atcagagtaa gtttaaactg agttctacta actaacgagt
     3901 aatatttaaa ttttcagcat ctcgcgcccg tgcctctgac ttctaagtcc aattactctt
     3961 caacatccct acatgctctt tctccctgtg ctcccacccc ctatttttgt tattatcaaa
     4021 aaacttctct taatttcttt gttttttagc ttcttttaag tcacctctaa caatgaaatt
     4081 gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc
     4141 cccccattaa taataattct atcccaaaat ctacacaatg ttctgtgtac acttcttatg
     4201 ttttttactt ctgataaatt tttttgaaac atcatagaaa aaaccgcaca caaaatacct
     4261 tatcatatgt tacgtttcag tttatgaccg caatttttat ttcttcgcac gtctgggcct
     4321 ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg agtatttttg gaatttttca
     4381 atcaagtgaa agtttatgaa attaattttc ctgcttttgc tttttggggt ttcccctatt
     4441 gtttgtcaag atttcgagga cggcgttttt cttgctaaaa tcacaagtat tgatgagcac
     4501 gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaaggt gagtagaagt
     4561 tgataatttg aaagtggagt agtgtctatg gggtttttgc cttaaatgac agaatacatt
     4621 cccaatatac caaacataac tgtttcctac tagtcggccg tacgggccct ttcgtctcgc
     4681 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4741 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4801 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4861 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta
     4921 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4981 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5041 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5101 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5161 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5221 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5281 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5341 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5401 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5461 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5521 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5581 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5641 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5701 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5761 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5821 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5881 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5941 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6001 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6061 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6121 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6181 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6241 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6301 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6361 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6421 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6481 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     6541 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6601 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6661 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6721 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6781 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6841 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6901 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6961 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     7021 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     7081 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc t
//
