LOCUS       sequence-90-expo        7229 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 7229)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7229)
  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_03
FEATURES             Location/Qualifiers
     source          1..7229
                     /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(256..278)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4669..4691)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4791..4810
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4888..4906)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4974..5078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5079..5147,5148..5939)
                     /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(5297..5316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6110..6698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6599..6618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6852..6869
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6986..7007
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7022..7027,7028..7045,7046..7052)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7060..7076
                     /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     7065..7087
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7084..7100
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7084..7100
                     /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 tgggtgaaga
      181 ccagaaacag cacctcgaac tgagccgcga tattgcccag cgtttcaacg cgctgtatgg
      241 cgagatcgat cccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      301 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      361 cgatcgccct tcccaacagt tgcgaaggta agtttaaaca gatccatact aactaacttg
      421 ttctgacata attttcagct tgaatggcga atggcgcttt gcctggtttc cggcaccaga
      481 agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc
      541 ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtga cctatcccat
      601 tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt
      661 taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttaa
      721 ctcggcgttt catctgtggt gcaacgggcg ctgggtcggt tacggccagg taagtttaat
      781 taagttgata ctaactaaca agatctgatt aattttcagg acagtcgttt gccgtctgaa
      841 tttgacctga gcgcattttt acgcgccgga gaaaaccgcc tcgcggtgat ggtgctgcgc
      901 tggagtgacg gcagttatct ggaagatcag gatatgtggc ggatgagcgg cattttccgt
      961 gacgtctcgt tgctgcataa accgactaca caaatcagcg atttccatgt tgccactcgc
     1021 tttaatgatg atttctcccg cgctgtactg gaggctgaag tccaggtaag tttaaacagg
     1081 atcttactaa ctaacatgct aacactgaat tttcagatgt gcggcgagtt gcgtgactac
     1141 ctacgggtaa cagtttcttt atggcagggt gaaacgcagg tcgccagcgg caccgcgcct
     1201 ttcggcggtg aaattatcga tgagcgtggt ggttatgccg atcgcgtcac actacgtctg
     1261 aacgtcgaaa acccgaaact gtggagcgcc gaaatcccga atctctatcg tgcggtggtt
     1321 gaactgcaca ccgccgacgg cacgctgatt gaagcagaag cctgcgatgt cggtttccgc
     1381 gaggtgcgga ttgaaaatgg tctgctgctg ctgaacggca agccgttgct gattcgaggc
     1441 gttaaccgtc acgagcatca tcctctgcat ggtcaggtca tggatgagca gacgatggtg
     1501 caggatgtaa gtttaaacta ttcgttacta actaacttta aacatttaaa ttttcagatc
     1561 ctgctgatga agcagaacaa ctttaacgcc gtgcgctgtt cgcattatcc gaaccatccg
     1621 ctgtggtaca cgctgtgcga ccgctacggc ctgtatgtgg tggatgaagc caatattgaa
     1681 acccacggca tggtgccaat gaatcgtctg accgatgatc cgcgctggct accggcgatg
     1741 agcgaacgcg taacgcgaat ggtgcagcgc gatcgtaatc acccgagtgt gatcatctgg
     1801 tcgctgggga atggtaagtt taaacagttg aatactaact aacggagatc tttgaaattt
     1861 tcagaatcag gccacggcgc taatcacgac gcgctgtatc gctggatcaa atctgtcgat
     1921 ccttcccgcc cggtgcagta tgaaggcggc ggagccgaca ccacggccac cgatattatt
     1981 tgcccgatgt acgcgcgcgt ggatgaagac cagcccttcc cggctgtgcc gaaatggtcc
     2041 atcaaaaaat ggctttcgct acctggagag acgcgcccgc tgattctttg cgaggtaagt
     2101 ttaaacagaa ctctactaac taacacatta gatcctaatt ttcagtacgc tcacgcgatg
     2161 ggcaacagtc ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta tccccgttta
     2221 cagggcggct tcgtctggga ctgggtggat cagtcgctga ttaaatatga tgaaaacggc
     2281 aacccgtggt cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg ccagttctgt
     2341 atgaacggtc tggtctttgc cgaccgcacg ccgcatccag cgtaagttta aacaataacc
     2401 tactaactaa cgtagataat ttaaattttc aggctgacgg aagcaaaaca ccagcagcag
     2461 tttttccagt tccgtttatc cgggcaaacc atcgaagtga ccagcgaata cctgttccgt
     2521 catagcgata acgagctcct gcactggatg gtggcgctgg atggtaagcc gctggcaagc
     2581 ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt tgattgaact gcctgaacta
     2641 ccgcagccgg agagcgccgg gcaactctgg ctcacagtac gcgtagtgca accgaacgcg
     2701 accgcatggt cagaagccgg gcacatcagc gcatggcagc agtggaggta agtttaaaca
     2761 agatcctact aactaactct acattgatga attttcagac tggcggaaaa cctcagtgtg
     2821 acgctccccg ccgcgtccca cgccatcccg catctgacca ccagcgaaat ggatttttgc
     2881 atcgagctgg gtaataagcg ttggcaattt aaccgccagt caggctttct ttcacagatg
     2941 tggattggcg ataaaaaaca actgctgacg ccgctgcgcg atcagttcac ccgtgcaccg
     3001 ctggataacg acattggcgt aagtgaagcg acccgcattg accctaacgc ctgggtcggt
     3061 aagtttaaac aaagttgtac taactaacga agatcttgat aattttcaga acgctggaag
     3121 gcggcgggcc attaccaggc cgaagcagcg ttgttgcagt gcacggcaga tacacttgct
     3181 gatgcggtgc tgattacgac cgctcacgcg tggcagcatc aggggaaaac cttatttatc
     3241 agccggaaaa cctaccggat tgatggtagt ggtcaaatgg cgattaccgt tgatgttgaa
     3301 gtggcgagcg atacaccgca tccggcgcgg attggcctga actgccagct ggcgcaggta
     3361 gcagagcggg taaactggct cggattaggg ccgcaagaaa actatcccga ccgccttact
     3421 gccgcctgtt ttgaccgctg ggatctgcca ttgtcagaca tgtagtaagt ttaaacttga
     3481 tagtactaac taacatgttt catttaaatt ttcagtaccc cgtacgtctt cccgagcgaa
     3541 aacggtctgc gctgcgggac gcgcgaattg aattatggcc cacaccagtg gcgcggcgac
     3601 ttccagttca acatcagccg ctacagtcaa cagcaactga tggaaaccag ccatcgccat
     3661 ctgctgcacg cggaagaagg cacatggctg aatatcgacg gtttccatat ggggattggt
     3721 ggcgacgact cctggagccc gtcagtatcg gcggaattcc aactgagcgc cggtcgctac
     3781 cattaccaac ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga gtaagtttaa
     3841 actgagttct actaactaac gagtaatatt taaattttca gcatctcgcg cccgtgcctc
     3901 tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca
     3961 ccccctattt ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt
     4021 taagtcacct ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa
     4081 agtcgaaaaa aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac
     4141 aatgttctgt gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata
     4201 gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt
     4261 ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt
     4321 ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt
     4381 ttgctttttg gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct
     4441 aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag
     4501 gctcagtgga aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt
     4561 ttgccttaaa tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg
     4621 gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4681 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4741 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4801 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     4861 aaaataccgc atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg
     4921 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4981 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5041 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5101 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5161 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5221 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5281 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5341 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5401 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5461 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5521 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5581 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5641 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5701 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5761 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5821 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5881 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5941 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6001 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6061 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6121 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6181 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6241 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6301 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6361 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6421 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6481 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6541 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6601 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6661 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6721 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6781 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6841 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     6901 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     6961 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     7021 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     7081 acacaggaaa cagctatgac catgattacg ccaagctgta agtttaaaca tgatcttact
     7141 aactaactat tctcatttaa attttcagag cttaaaaatg gctgaaatca ctcacaacga
     7201 tggatacgct aacaacttgg aaatgaaat
//
