LOCUS       sequence-75-expo        7111 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 7111)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7111)
  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: pPD89_09
FEATURES             Location/Qualifiers
     source          1..7111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             162..182
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(272..294)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4685..4707)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4807..4826
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4904..4922)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4990..5094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5095..5163,5164..5955)
                     /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(5313..5332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6126..6714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6615..6634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6868..6885
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7002..7023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7038..7043,7044..7061,7062..7068)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7076..7092
                     /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     7081..7103
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7100..7111,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7100..7111,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggat
       61 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      121 gaggaccctt gagggtaccg agctcagaaa aaatgactgc tccaaagaag aagcgtaagg
      181 taccggtggg tgaagaccag aaacagcacc tcgaactgag ccgcgatatt gcccagcgtt
      241 tcaacgcgct gtatggcgag atcgatcccg tcgttttaca acgtcgtgac tgggaaaacc
      301 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      361 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg aaggtaagtt taaacagatc
      421 catactaact aacttgttct gacataattt tcagcttgaa tggcgaatgg cgctttgcct
      481 ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg
      541 atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca
      601 acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt
      661 gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta
      721 tttttgatgg cgttaactcg gcgtttcatc tgtggtgcaa cgggcgctgg gtcggttacg
      781 gccaggtaag tttaattaag ttgatactaa ctaacaagat ctgattaatt ttcaggacag
      841 tcgtttgccg tctgaatttg acctgagcgc atttttacgc gccggagaaa accgcctcgc
      901 ggtgatggtg ctgcgctgga gtgacggcag ttatctggaa gatcaggata tgtggcggat
      961 gagcggcatt ttccgtgacg tctcgttgct gcataaaccg actacacaaa tcagcgattt
     1021 ccatgttgcc actcgcttta atgatgattt ctcccgcgct gtactggagg ctgaagtcca
     1081 ggtaagttta aacaggatct tactaactaa catgctaaca ctgaattttc agatgtgcgg
     1141 cgagttgcgt gactacctac gggtaacagt ttctttatgg cagggtgaaa cgcaggtcgc
     1201 cagcggcacc gcgcctttcg gcggtgaaat tatcgatgag cgtggtggtt atgccgatcg
     1261 cgtcacacta cgtctgaacg tcgaaaaccc gaaactgtgg agcgccgaaa tcccgaatct
     1321 ctatcgtgcg gtggttgaac tgcacaccgc cgacggcacg ctgattgaag cagaagcctg
     1381 cgatgtcggt ttccgcgagg tgcggattga aaatggtctg ctgctgctga acggcaagcc
     1441 gttgctgatt cgaggcgtta accgtcacga gcatcatcct ctgcatggtc aggtcatgga
     1501 tgagcagacg atggtgcagg atgtaagttt aaactattcg ttactaacta actttaaaca
     1561 tttaaatttt cagatcctgc tgatgaagca gaacaacttt aacgccgtgc gctgttcgca
     1621 ttatccgaac catccgctgt ggtacacgct gtgcgaccgc tacggcctgt atgtggtgga
     1681 tgaagccaat attgaaaccc acggcatggt gccaatgaat cgtctgaccg atgatccgcg
     1741 ctggctaccg gcgatgagcg aacgcgtaac gcgaatggtg cagcgcgatc gtaatcaccc
     1801 gagtgtgatc atctggtcgc tggggaatgg taagtttaaa cagttgaata ctaactaacg
     1861 gagatctttg aaattttcag aatcaggcca cggcgctaat cacgacgcgc tgtatcgctg
     1921 gatcaaatct gtcgatcctt cccgcccggt gcagtatgaa ggcggcggag ccgacaccac
     1981 ggccaccgat attatttgcc cgatgtacgc gcgcgtggat gaagaccagc ccttcccggc
     2041 tgtgccgaaa tggtccatca aaaaatggct ttcgctacct ggagagacgc gcccgctgat
     2101 tctttgcgag gtaagtttaa acagaactct actaactaac acattagatc ctaattttca
     2161 gtacgctcac gcgatgggca acagtcttgg cggtttcgct aaatactggc aggcgtttcg
     2221 tcagtatccc cgtttacagg gcggcttcgt ctgggactgg gtggatcagt cgctgattaa
     2281 atatgatgaa aacggcaacc cgtggtcggc ttacggcggt gattttggcg atacgccgaa
     2341 cgatcgccag ttctgtatga acggtctggt ctttgccgac cgcacgccgc atccagcgta
     2401 agtttaaaca ataacctact aactaacgta gataatttaa attttcaggc tgacggaagc
     2461 aaaacaccag cagcagtttt tccagttccg tttatccggg caaaccatcg aagtgaccag
     2521 cgaatacctg ttccgtcata gcgataacga gctcctgcac tggatggtgg cgctggatgg
     2581 taagccgctg gcaagcggtg aagtgcctct ggatgtcgct ccacaaggta aacagttgat
     2641 tgaactgcct gaactaccgc agccggagag cgccgggcaa ctctggctca cagtacgcgt
     2701 agtgcaaccg aacgcgaccg catggtcaga agccgggcac atcagcgcat ggcagcagtg
     2761 gaggtaagtt taaacaagat cctactaact aactctacat tgatgaattt tcagactggc
     2821 ggaaaacctc agtgtgacgc tccccgccgc gtcccacgcc atcccgcatc tgaccaccag
     2881 cgaaatggat ttttgcatcg agctgggtaa taagcgttgg caatttaacc gccagtcagg
     2941 ctttctttca cagatgtgga ttggcgataa aaaacaactg ctgacgccgc tgcgcgatca
     3001 gttcacccgt gcaccgctgg ataacgacat tggcgtaagt gaagcgaccc gcattgaccc
     3061 taacgcctgg gtcggtaagt ttaaacaaag ttgtactaac taacgaagat cttgataatt
     3121 ttcagaacgc tggaaggcgg cgggccatta ccaggccgaa gcagcgttgt tgcagtgcac
     3181 ggcagataca cttgctgatg cggtgctgat tacgaccgct cacgcgtggc agcatcaggg
     3241 gaaaacctta tttatcagcc ggaaaaccta ccggattgat ggtagtggtc aaatggcgat
     3301 taccgttgat gttgaagtgg cgagcgatac accgcatccg gcgcggattg gcctgaactg
     3361 ccagctggcg caggtagcag agcgggtaaa ctggctcgga ttagggccgc aagaaaacta
     3421 tcccgaccgc cttactgccg cctgttttga ccgctgggat ctgccattgt cagacatgta
     3481 gtaagtttaa acttgatagt actaactaac atgtttcatt taaattttca gtaccccgta
     3541 cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc gaattgaatt atggcccaca
     3601 ccagtggcgc ggcgacttcc agttcaacat cagccgctac agtcaacagc aactgatgga
     3661 aaccagccat cgccatctgc tgcacgcgga agaaggcaca tggctgaata tcgacggttt
     3721 ccatatgggg attggtggcg acgactcctg gagcccgtca gtatcggcgg aattccaact
     3781 gagcgccggt cgctaccatt accaacttgt ctggtgtcaa aaataatagg ggccgctgtc
     3841 atcagagtaa gtttaaactg agttctacta actaacgagt aatatttaaa ttttcagcat
     3901 ctcgcgcccg tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt
     3961 tctccctgtg ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt
     4021 gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa
     4081 ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct
     4141 atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt
     4201 tttttgaaac atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag
     4261 tttatgaccg caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc
     4321 tcatcgtgaa aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa
     4381 attaattttc ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga
     4441 cggcgttttt cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag
     4501 aaggtttggg tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt
     4561 agtgtctatg gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac
     4621 tgtttcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg
     4681 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4741 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     4801 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     4861 acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta
     4921 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4981 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5041 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5101 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5161 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5221 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5281 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5341 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5401 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5461 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5521 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5581 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5641 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5701 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5761 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5821 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5881 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5941 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6001 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6061 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6121 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6181 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6241 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     6301 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6361 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6421 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6481 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     6541 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6601 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6661 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6721 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6781 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6841 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6901 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6961 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7021 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7081 agcggataac aatttcacac aggaaacagc t
//
