LOCUS       sequence-82-expo        6908 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 6908)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6908)
  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: pPD90_31
FEATURES             Location/Qualifiers
     source          1..6908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             163..183
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(273..295)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4482..4504)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4604..4623
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4701..4719)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4787..4891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4892..4960,4961..5752)
                     /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(5110..5129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5923..6511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6412..6431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6665..6682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6799..6820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6835..6840,6841..6858,6859..6865)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6873..6889
                     /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     6878..6900
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6897..6908,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6897..6908,1..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 ggagggtacc gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag
      181 gtaccggtgg gtgaagacca gaaacagcac ctcgaactga gccgcgatat tgcccagcgt
      241 ttcaacgcgc tgtatggcga gatcgatccc gtcgttttac aacgtcgtga ctgggaaaac
      301 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      361 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      421 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt
      481 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc
      541 atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat
      601 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag
      661 acgcgaatta tttttgatgg cgttaactcg gcgtttcatc tgtggtgcaa cgggcgctgg
      721 gtcggttacg gccaggtaag tttaattaag ttgatactaa ctaacaagat ctgattaatt
      781 ttcaggacag tcgtttgccg tctgaatttg acctgagcgc atttttacgc gccggagaaa
      841 accgcctcgc ggtgatggtg ctgcgctgga gtgacggcag ttatctggaa gatcaggata
      901 tgtggcggat gagcggcatt ttccgtgacg tctcgttgct gcataaaccg actacacaaa
      961 tcagcgattt ccatgttgcc actcgcttta atgatgattt cagccgcgct gtactggagg
     1021 ctgaagttca gatgtgcggc gagttgcgtg actacctacg ggtaacagtt tctttatggc
     1081 agggtgaaac gcaggtcgcc agcggcaccg cgcctttcgg cggtgaaatt atcgatgagc
     1141 gtggtggtta tgccgatcgc gtcacactac gtctgaacgt cgaaaacccg aaactgtgga
     1201 gcgccgaaat cccgaatctc tatcgtgcgg tggttgaact gcacaccgcc gacggcacgc
     1261 tgattgaagc agaagcctgc gatgtcggtt tccgcgaggt gcggattgaa aatggtctgc
     1321 tgctgctgaa cggcaagccg ttgctgattc gaggcgttaa ccgtcacgag catcatcctc
     1381 tgcatggtca ggtcatggat gagcagacga tggtgcagga tgtaagttta aactattcgt
     1441 tactaactaa ctttaaacat ttaaattttc agatcctgct gatgaagcag aacaacttta
     1501 acgccgtgcg ctgttcgcat tatccgaacc atccgctgtg gtacacgctg tgcgaccgct
     1561 acggcctgta tgtggtggat gaagccaata ttgaaaccca cggcatggtg ccaatgaatc
     1621 gtctgaccga tgatccgcgc tggctaccgg cgatgagcga acgcgtaacg cgaatggtgc
     1681 agcgcgatcg taatcacccg agtgtgatca tctggtcgct ggggaatggt aagtttaaac
     1741 agttgaatac taactaacgg agatctttga aattttcaga atcaggccac ggcgctaatc
     1801 acgacgcgct gtatcgctgg atcaaatctg tcgatccttc ccgcccggtg cagtatgaag
     1861 gcggcggagc cgacaccacg gccaccgata ttatttgccc gatgtacgcg cgcgtggatg
     1921 aagaccagcc cttcccggct gtgccgaaat ggtccatcaa aaaatggctt tcgctacctg
     1981 gagagacgcg cccgctgatc ctttgcgaat acgcccacgc gatgggtaac agtcttggcg
     2041 gtttcgctaa atactggcag gcgtttcgtc agtatccccg tttacagggc ggcttcgtct
     2101 gggactgggt ggatcagtcg ctgattaaat atgatgaaaa cggcaacccg tggtcggctt
     2161 acggcggtga ttttggcgat acgccgaacg atcgccagtt ctgtatgaac ggtctggtct
     2221 ttgccgaccg cacgccgcat ccagcgtaag tttaaacaat aacctactaa ctaacgtaga
     2281 taatttaaat tttcaggctg acggaagcaa aacaccagca gcagtttttc cagttccgtt
     2341 tatccgggca aaccatcgaa gtgaccagcg aatacctgtt ccgtcatagc gataacgagc
     2401 tcctgcactg gatggtggcg ctggatggta agccgctggc aagcggtgaa gtgcctctgg
     2461 atgtcgctcc acaaggtaaa cagttgattg aactgcctga actaccgcag ccggagagcg
     2521 ccgggcaact ctggctcaca gtacgcgtag tgcaaccgaa cgcgaccgca tggtcagaag
     2581 ccgggcacat cagcgcctgg cagcagtggc gtctggcgga aaacctcagt gtgacgctcc
     2641 ccgccgcgtc ccacgccatc ccgcatctga ccaccagcga aatggatttt tgcatcgagc
     2701 tgggtaataa gcgttggcaa tttaaccgcc agtcaggctt tctttcacag atgtggattg
     2761 gcgataaaaa acaactgctg acgccgctgc gcgatcagtt cacccgtgca ccgctggata
     2821 acgacattgg cgtaagtgaa gcgacccgca ttgaccctaa cgcctgggtc ggtaagttta
     2881 aacaaagttg tactaactaa cgaagatctt gataattttc agaacgctgg aaggcggcgg
     2941 gccattacca ggccgaagca gcgttgttgc agtgcacggc agatacactt gctgatgcgg
     3001 tgctgattac gaccgctcac gcgtggcagc atcaggggaa aaccttattt atcagccgga
     3061 aaacctaccg gattgatggt agtggtcaaa tggcgattac cgttgatgtt gaagtggcga
     3121 gcgatacacc gcatccggcg cggattggcc tgaactgcca gctggcgcag gtagcagagc
     3181 gggtaaactg gctcggatta gggccgcaag aaaactatcc cgaccgcctt actgccgcct
     3241 gttttgaccg ctgggatctg ccattgtcag acatgtagta agtttaaact tgatagtact
     3301 aactaacatg tttcatttaa attttcagta ccccgtacgt cttcccgagc gaaaacggtc
     3361 tgcgctgcgg gacgcgcgaa ttgaattatg gcccacacca gtggcgcggc gacttccagt
     3421 tcaacatcag ccgctacagt caacagcaac tgatggaaac cagccatcgc catctgctgc
     3481 acgcggaaga aggcacatgg ctgaatatcg acggtttcca tatggggatt ggtggcgacg
     3541 actcctggag cccgtcagta tcggcggaat tccaactgag cgccggtcgc taccattacc
     3601 aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc agagtaagtt taaactgagt
     3661 tctactaact aacgagtaat atttaaattt tcagcatctc gcgcccgtgc ctctgacttc
     3721 taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc ccacccccta
     3781 tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc ttttaagtca
     3841 cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa
     3901 aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc
     3961 tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc atagaaaaaa
     4021 ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc
     4081 ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt
     4141 atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt
     4201 ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca
     4261 caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt gaggctcagt
     4321 ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg tttttgcctt
     4381 aaatgacaga atacattccc aatataccaa acataactgt ttcctactag tcggccgtac
     4441 gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     4501 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     4561 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     4621 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     4681 cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4741 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4801 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4861 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4921 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4981 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5041 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5101 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5161 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5221 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5281 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5341 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5401 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5461 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5521 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5581 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5641 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5701 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5761 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5821 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6061 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6301 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6541 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6601 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6661 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     6721 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6781 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     6841 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6901 aaacagct
//
