LOCUS       sequence-76-expo        7116 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 7116)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7116)
  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_17
FEATURES             Location/Qualifiers
     source          1..7116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             167..187
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(277..299)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4690..4712)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4812..4831
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4909..4927)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4995..5099
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5100..5168,5169..5960)
                     /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(5318..5337)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6131..6719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6620..6639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6873..6890
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7007..7028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7043..7048,7049..7066,7067..7073)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7081..7097
                     /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     7086..7108
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7105..7116,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7105..7116,1..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 gcttggaggg taccgagctc agaaaaaatg actgctccaa agaagaagcg
      181 taaggtaccg gtgggtgaag accagaaaca gcacctcgaa ctgagccgcg atattgccca
      241 gcgtttcaac gcgctgtatg gcgagatcga tcccgtcgtt ttacaacgtc gtgactggga
      301 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
      361 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgaaggt aagtttaaac
      421 agatccatac taactaactt gttctgacat aattttcagc ttgaatggcg aatggcgctt
      481 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga
      541 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
      601 caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
      661 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
      721 aattattttt gatggcgtta actcggcgtt tcatctgtgg tgcaacgggc gctgggtcgg
      781 ttacggccag gtaagtttaa ttaagttgat actaactaac aagatctgat taattttcag
      841 gacagtcgtt tgccgtctga atttgacctg agcgcatttt tacgcgccgg agaaaaccgc
      901 ctcgcggtga tggtgctgcg ctggagtgac ggcagttatc tggaagatca ggatatgtgg
      961 cggatgagcg gcattttccg tgacgtctcg ttgctgcata aaccgactac acaaatcagc
     1021 gatttccatg ttgccactcg ctttaatgat gatttctccc gcgctgtact ggaggctgaa
     1081 gtccaggtaa gtttaaacag gatcttacta actaacatgc taacactgaa ttttcagatg
     1141 tgcggcgagt tgcgtgacta cctacgggta acagtttctt tatggcaggg tgaaacgcag
     1201 gtcgccagcg gcaccgcgcc tttcggcggt gaaattatcg atgagcgtgg tggttatgcc
     1261 gatcgcgtca cactacgtct gaacgtcgaa aacccgaaac tgtggagcgc cgaaatcccg
     1321 aatctctatc gtgcggtggt tgaactgcac accgccgacg gcacgctgat tgaagcagaa
     1381 gcctgcgatg tcggtttccg cgaggtgcgg attgaaaatg gtctgctgct gctgaacggc
     1441 aagccgttgc tgattcgagg cgttaaccgt cacgagcatc atcctctgca tggtcaggtc
     1501 atggatgagc agacgatggt gcaggatgta agtttaaact attcgttact aactaacttt
     1561 aaacatttaa attttcagat cctgctgatg aagcagaaca actttaacgc cgtgcgctgt
     1621 tcgcattatc cgaaccatcc gctgtggtac acgctgtgcg accgctacgg cctgtatgtg
     1681 gtggatgaag ccaatattga aacccacggc atggtgccaa tgaatcgtct gaccgatgat
     1741 ccgcgctggc taccggcgat gagcgaacgc gtaacgcgaa tggtgcagcg cgatcgtaat
     1801 cacccgagtg tgatcatctg gtcgctgggg aatggtaagt ttaaacagtt gaatactaac
     1861 taacggagat ctttgaaatt ttcagaatca ggccacggcg ctaatcacga cgcgctgtat
     1921 cgctggatca aatctgtcga tccttcccgc ccggtgcagt atgaaggcgg cggagccgac
     1981 accacggcca ccgatattat ttgcccgatg tacgcgcgcg tggatgaaga ccagcccttc
     2041 ccggctgtgc cgaaatggtc catcaaaaaa tggctttcgc tacctggaga gacgcgcccg
     2101 ctgattcttt gcgaggtaag tttaaacaga actctactaa ctaacacatt agatcctaat
     2161 tttcagtacg ctcacgcgat gggcaacagt cttggcggtt tcgctaaata ctggcaggcg
     2221 tttcgtcagt atccccgttt acagggcggc ttcgtctggg actgggtgga tcagtcgctg
     2281 attaaatatg atgaaaacgg caacccgtgg tcggcttacg gcggtgattt tggcgatacg
     2341 ccgaacgatc gccagttctg tatgaacggt ctggtctttg ccgaccgcac gccgcatcca
     2401 gcgtaagttt aaacaataac ctactaacta acgtagataa tttaaatttt caggctgacg
     2461 gaagcaaaac accagcagca gtttttccag ttccgtttat ccgggcaaac catcgaagtg
     2521 accagcgaat acctgttccg tcatagcgat aacgagctcc tgcactggat ggtggcgctg
     2581 gatggtaagc cgctggcaag cggtgaagtg cctctggatg tcgctccaca aggtaaacag
     2641 ttgattgaac tgcctgaact accgcagccg gagagcgccg ggcaactctg gctcacagta
     2701 cgcgtagtgc aaccgaacgc gaccgcatgg tcagaagccg ggcacatcag cgcatggcag
     2761 cagtggaggt aagtttaaac aagatcctac taactaactc tacattgatg aattttcaga
     2821 ctggcggaaa acctcagtgt gacgctcccc gccgcgtccc acgccatccc gcatctgacc
     2881 accagcgaaa tggatttttg catcgagctg ggtaataagc gttggcaatt taaccgccag
     2941 tcaggctttc tttcacagat gtggattggc gataaaaaac aactgctgac gccgctgcgc
     3001 gatcagttca cccgtgcacc gctggataac gacattggcg taagtgaagc gacccgcatt
     3061 gaccctaacg cctgggtcgg taagtttaaa caaagttgta ctaactaacg aagatcttga
     3121 taattttcag aacgctggaa ggcggcgggc cattaccagg ccgaagcagc gttgttgcag
     3181 tgcacggcag atacacttgc tgatgcggtg ctgattacga ccgctcacgc gtggcagcat
     3241 caggggaaaa ccttatttat cagccggaaa acctaccgga ttgatggtag tggtcaaatg
     3301 gcgattaccg ttgatgttga agtggcgagc gatacaccgc atccggcgcg gattggcctg
     3361 aactgccagc tggcgcaggt agcagagcgg gtaaactggc tcggattagg gccgcaagaa
     3421 aactatcccg accgccttac tgccgcctgt tttgaccgct gggatctgcc attgtcagac
     3481 atgtagtaag tttaaacttg atagtactaa ctaacatgtt tcatttaaat tttcagtacc
     3541 ccgtacgtct tcccgagcga aaacggtctg cgctgcggga cgcgcgaatt gaattatggc
     3601 ccacaccagt ggcgcggcga cttccagttc aacatcagcc gctacagtca acagcaactg
     3661 atggaaacca gccatcgcca tctgctgcac gcggaagaag gcacatggct gaatatcgac
     3721 ggtttccata tggggattgg tggcgacgac tcctggagcc cgtcagtatc ggcggaattc
     3781 caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata ataggggccg
     3841 ctgtcatcag agtaagttta aactgagttc tactaactaa cgagtaatat ttaaattttc
     3901 agcatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca tccctacatg
     3961 ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact tctcttaatt
     4021 tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa
     4081 tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc attaataata
     4141 attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt tacttctgat
     4201 aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca tatgttacgt
     4261 ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat gacgtcaaat
     4321 catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa gtgaaagttt
     4381 atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg tcaagatttc
     4441 gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc aagaaagatc
     4501 ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata atttgaaagt
     4561 ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa tataccaaac
     4621 ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga
     4681 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4741 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4801 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     4861 accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc ctcgtgatac
     4921 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4981 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5041 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5101 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5161 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5221 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5281 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5341 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5401 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5461 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5521 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5581 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5641 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5701 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5761 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5821 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5881 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5941 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6001 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6061 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6241 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     6541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6661 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6721 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6781 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6841 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     6901 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     6961 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7021 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7081 ttgtgagcgg ataacaattt cacacaggaa acagct
//
