LOCUS       sequence-74-expo        7112 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 7112)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7112)
  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_03
FEATURES             Location/Qualifiers
     source          1..7112
                     /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(4686..4708)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4808..4827
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4905..4923)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4991..5095
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5096..5164,5165..5956)
                     /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(5314..5333)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6127..6715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6616..6635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6869..6886
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7003..7024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7039..7044,7045..7062,7063..7069)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7077..7093
                     /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     7082..7104
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7101..7112,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7101..7112,1..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 gaaggtaagt ttaaacagat
      421 ccatactaac taacttgttc tgacataatt ttcagcttga atggcgaatg gcgctttgcc
      481 tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc
      541 gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc
      601 aacgtgacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt
      661 tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt
      721 atttttgatg gcgttaactc ggcgtttcat ctgtggtgca acgggcgctg ggtcggttac
      781 ggccaggtaa gtttaattaa gttgatacta actaacaaga tctgattaat tttcaggaca
      841 gtcgtttgcc gtctgaattt gacctgagcg catttttacg cgccggagaa aaccgcctcg
      901 cggtgatggt gctgcgctgg agtgacggca gttatctgga agatcaggat atgtggcgga
      961 tgagcggcat tttccgtgac gtctcgttgc tgcataaacc gactacacaa atcagcgatt
     1021 tccatgttgc cactcgcttt aatgatgatt tctcccgcgc tgtactggag gctgaagtcc
     1081 aggtaagttt aaacaggatc ttactaacta acatgctaac actgaatttt cagatgtgcg
     1141 gcgagttgcg tgactaccta cgggtaacag tttctttatg gcagggtgaa acgcaggtcg
     1201 ccagcggcac cgcgcctttc ggcggtgaaa ttatcgatga gcgtggtggt tatgccgatc
     1261 gcgtcacact acgtctgaac gtcgaaaacc cgaaactgtg gagcgccgaa atcccgaatc
     1321 tctatcgtgc ggtggttgaa ctgcacaccg ccgacggcac gctgattgaa gcagaagcct
     1381 gcgatgtcgg tttccgcgag gtgcggattg aaaatggtct gctgctgctg aacggcaagc
     1441 cgttgctgat tcgaggcgtt aaccgtcacg agcatcatcc tctgcatggt caggtcatgg
     1501 atgagcagac gatggtgcag gatgtaagtt taaactattc gttactaact aactttaaac
     1561 atttaaattt tcagatcctg ctgatgaagc agaacaactt taacgccgtg cgctgttcgc
     1621 attatccgaa ccatccgctg tggtacacgc tgtgcgaccg ctacggcctg tatgtggtgg
     1681 atgaagccaa tattgaaacc cacggcatgg tgccaatgaa tcgtctgacc gatgatccgc
     1741 gctggctacc ggcgatgagc gaacgcgtaa cgcgaatggt gcagcgcgat cgtaatcacc
     1801 cgagtgtgat catctggtcg ctggggaatg gtaagtttaa acagttgaat actaactaac
     1861 ggagatcttt gaaattttca gaatcaggcc acggcgctaa tcacgacgcg ctgtatcgct
     1921 ggatcaaatc tgtcgatcct tcccgcccgg tgcagtatga aggcggcgga gccgacacca
     1981 cggccaccga tattatttgc ccgatgtacg cgcgcgtgga tgaagaccag cccttcccgg
     2041 ctgtgccgaa atggtccatc aaaaaatggc tttcgctacc tggagagacg cgcccgctga
     2101 ttctttgcga ggtaagttta aacagaactc tactaactaa cacattagat cctaattttc
     2161 agtacgctca cgcgatgggc aacagtcttg gcggtttcgc taaatactgg caggcgtttc
     2221 gtcagtatcc ccgtttacag ggcggcttcg tctgggactg ggtggatcag tcgctgatta
     2281 aatatgatga aaacggcaac ccgtggtcgg cttacggcgg tgattttggc gatacgccga
     2341 acgatcgcca gttctgtatg aacggtctgg tctttgccga ccgcacgccg catccagcgt
     2401 aagtttaaac aataacctac taactaacgt agataattta aattttcagg ctgacggaag
     2461 caaaacacca gcagcagttt ttccagttcc gtttatccgg gcaaaccatc gaagtgacca
     2521 gcgaatacct gttccgtcat agcgataacg agctcctgca ctggatggtg gcgctggatg
     2581 gtaagccgct ggcaagcggt gaagtgcctc tggatgtcgc tccacaaggt aaacagttga
     2641 ttgaactgcc tgaactaccg cagccggaga gcgccgggca actctggctc acagtacgcg
     2701 tagtgcaacc gaacgcgacc gcatggtcag aagccgggca catcagcgca tggcagcagt
     2761 ggaggtaagt ttaaacaaga tcctactaac taactctaca ttgatgaatt ttcagactgg
     2821 cggaaaacct cagtgtgacg ctccccgccg cgtcccacgc catcccgcat ctgaccacca
     2881 gcgaaatgga tttttgcatc gagctgggta ataagcgttg gcaatttaac cgccagtcag
     2941 gctttctttc acagatgtgg attggcgata aaaaacaact gctgacgccg ctgcgcgatc
     3001 agttcacccg tgcaccgctg gataacgaca ttggcgtaag tgaagcgacc cgcattgacc
     3061 ctaacgcctg ggtcggtaag tttaaacaaa gttgtactaa ctaacgaaga tcttgataat
     3121 tttcagaacg ctggaaggcg gcgggccatt accaggccga agcagcgttg ttgcagtgca
     3181 cggcagatac acttgctgat gcggtgctga ttacgaccgc tcacgcgtgg cagcatcagg
     3241 ggaaaacctt atttatcagc cggaaaacct accggattga tggtagtggt caaatggcga
     3301 ttaccgttga tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt ggcctgaact
     3361 gccagctggc gcaggtagca gagcgggtaa actggctcgg attagggccg caagaaaact
     3421 atcccgaccg ccttactgcc gcctgttttg accgctggga tctgccattg tcagacatgt
     3481 agtaagttta aacttgatag tactaactaa catgtttcat ttaaattttc agtaccccgt
     3541 acgtcttccc gagcgaaaac ggtctgcgct gcgggacgcg cgaattgaat tatggcccac
     3601 accagtggcg cggcgacttc cagttcaaca tcagccgcta cagtcaacag caactgatgg
     3661 aaaccagcca tcgccatctg ctgcacgcgg aagaaggcac atggctgaat atcgacggtt
     3721 tccatatggg gattggtggc gacgactcct ggagcccgtc agtatcggcg gaattccaac
     3781 tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag gggccgctgt
     3841 catcagagta agtttaaact gagttctact aactaacgag taatatttaa attttcagca
     3901 tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct
     3961 ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt
     4021 tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga
     4081 attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc
     4141 tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat
     4201 ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca
     4261 gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg
     4321 ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga
     4381 aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg
     4441 acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa
     4501 gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag
     4561 tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa
     4621 ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     4681 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     4741 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     4801 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     4861 cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct
     4921 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4981 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5041 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5101 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5161 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5221 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5281 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5341 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5401 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5461 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5521 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5581 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5641 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5701 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5761 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5821 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5881 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5941 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6001 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6061 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6121 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6181 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6241 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6301 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6361 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6421 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6481 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     6541 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6601 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6661 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6721 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6781 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6841 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     6901 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     6961 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     7021 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     7081 gagcggataa caatttcaca caggaaacag ct
//
