LOCUS       sequence-70-expo        6932 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 6932)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6932)
  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: pPD87_50
FEATURES             Location/Qualifiers
     source          1..6932
                     /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(4506..4528)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4628..4647
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4725..4743)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4811..4915
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4916..4984,4985..5776)
                     /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(5134..5153)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5947..6535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6436..6455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6689..6706
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6823..6844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6859..6864,6865..6882,6883..6889)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6897..6913
                     /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     6902..6924
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6921..6932,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6921..6932,1..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 acggccagga
      961 cagtcgtttg ccgtctgaat ttgacctgag cgcattttta cgcgccggag aaaaccgcct
     1021 cgcggtgatg gtgctgcgct ggagtgacgg cagttatctg gaagatcagg atatgtggcg
     1081 gatgagcggc attttccgtg acgtctcgtt gctgcataaa ccgactacac aaatcagcga
     1141 tttccatgtt gccactcgct ttaatgatga tttcagccgc gctgtactgg aggctgaagt
     1201 tcagatgtgc ggcgagttgc gtgactacct acgggtaaca gtttctttat ggcagggtga
     1261 aacgcaggtc gccagcggca ccgcgccttt cggcggtgaa attatcgatg agcgtggtgg
     1321 ttatgccgat cgcgtcacac tacgtctgaa cgtcgaaaac ccgaaactgt ggagcgccga
     1381 aatcccgaat ctctatcgtg cggtggttga actgcacacc gccgacggca cgctgattga
     1441 agcagaagcc tgcgatgtcg gtttccgcga ggtgcggatt gaaaatggtc tgctgctgct
     1501 gaacggcaag ccgttgctga ttcgaggcgt taaccgtcac gagcatcatc ctctgcatgg
     1561 tcaggtcatg gatgagcaga cgatggtgca ggatgtaagt ttaaactatt cgttactaac
     1621 taactttaaa catttaaatt ttcagatcct gctgatgaag cagaacaact ttaacgccgt
     1681 gcgctgttcg cattatccga accatccgct gtggtacacg ctgtgcgacc gctacggcct
     1741 gtatgtggtg gatgaagcca atattgaaac ccacggcatg gtgccaatga atcgtctgac
     1801 cgatgatccg cgctggctac cggcgatgag cgaacgcgta acgcgaatgg tgcagcgcga
     1861 tcgtaatcac ccgagtgtga tcatctggtc gctggggaat gaatcaggcc acggcgctaa
     1921 tcacgacgcg ctgtatcgct ggatcaaatc tgtcgatcct tcccgcccgg tgcagtatga
     1981 aggcggcgga gccgacacca cggccaccga tattatttgc ccgatgtacg cgcgcgtgga
     2041 tgaagaccag cccttcccgg ctgtgccgaa atggtccatc aaaaaatggc tttcgctacc
     2101 tggagagacg cgcccgctga tcctttgcga atacgcccac gcgatgggta acagtcttgg
     2161 cggtttcgct aaatactggc aggcgtttcg tcagtatccc cgtttacagg gcggcttcgt
     2221 ctgggactgg gtggatcagt cgctgattaa atatgatgaa aacggcaacc cgtggtcggc
     2281 ttacggcggt gattttggcg atacgccgaa cgatcgccag ttctgtatga acggtctggt
     2341 ctttgccgac cgcacgccgc atccagcgta agtttaaaca ataacctact aactaacgta
     2401 gataatttaa attttcaggc tgacggaagc aaaacaccag cagcagtttt tccagttccg
     2461 tttatccggg caaaccatcg aagtgaccag cgaatacctg ttccgtcata gcgataacga
     2521 gctcctgcac tggatggtgg cgctggatgg taagccgctg gcaagcggtg aagtgcctct
     2581 ggatgtcgct ccacaaggta aacagttgat tgaactgcct gaactaccgc agccggagag
     2641 cgccgggcaa ctctggctca cagtacgcgt agtgcaaccg aacgcgaccg catggtcaga
     2701 agccgggcac atcagcgcct ggcagcagtg gcgtctggcg gaaaacctca gtgtgacgct
     2761 ccccgccgcg tcccacgcca tcccgcatct gaccaccagc gaaatggatt tttgcatcga
     2821 gctgggtaat aagcgttggc aatttaaccg ccagtcaggc tttctttcac agatgtggat
     2881 tggcgataaa aaacaactgc tgacgccgct gcgcgatcag ttcacccgtg caccgctgga
     2941 taacgacatt ggcgtaagtg aagcgacccg cattgaccct aacgcctggg tcgaacgctg
     3001 gaaggcggcg ggccattacc aggccgaagc agcgttgttg cagtgcacgg cagatacact
     3061 tgctgatgcg gtgctgatta cgaccgctca cgcgtggcag catcagggga aaaccttatt
     3121 tatcagccgg aaaacctacc ggattgatgg tagtggtcaa atggcgatta ccgttgatgt
     3181 tgaagtggcg agcgatacac cgcatccggc gcggattggc ctgaactgcc agctggcgca
     3241 ggtagcagag cgggtaaact ggctcggatt agggccgcaa gaaaactatc ccgaccgcct
     3301 tactgccgcc tgttttgacc gctgggatct gccattgtca gacatgtagt aagtttaaac
     3361 ttgatagtac taactaacat gtttcattta aattttcagt accccgtacg tcttcccgag
     3421 cgaaaacggt ctgcgctgcg ggacgcgcga attgaattat ggcccacacc agtggcgcgg
     3481 cgacttccag ttcaacatca gccgctacag tcaacagcaa ctgatggaaa ccagccatcg
     3541 ccatctgctg cacgcggaag aaggcacatg gctgaatatc gacggtttcc atatggggat
     3601 tggtggcgac gactcctgga gcccgtcagt atcggcggaa ttccaactga gcgccggtcg
     3661 ctaccattac caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat cagatcgcca
     3721 tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct
     3781 ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt
     3841 tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga
     3901 attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc
     3961 tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat
     4021 ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca
     4081 gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg
     4141 ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga
     4201 aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg
     4261 acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa
     4321 gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag
     4381 tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa
     4441 ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     4501 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     4561 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     4621 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     4681 cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct
     4741 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4801 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4861 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4921 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4981 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5041 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5101 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5161 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5221 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5281 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5341 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5401 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5461 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5521 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5581 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5641 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5701 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5761 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5821 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5881 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5941 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6001 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6061 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6121 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6181 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6241 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6301 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     6361 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6421 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6481 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6541 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6601 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6661 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6721 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     6781 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     6841 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     6901 gagcggataa caatttcaca caggaaacag ct
//
