LOCUS       sequence-81-expo        6709 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 6709)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6709)
  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_28
FEATURES             Location/Qualifiers
     source          1..6709
                     /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(4283..4305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4405..4424
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4502..4520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4588..4692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4693..4761,4762..5553)
                     /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(4911..4930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5724..6312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6213..6232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6466..6483
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6600..6621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6636..6641,6642..6659,6660..6666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6674..6690
                     /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     6679..6701
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6698..6709,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6698..6709,1..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 gccaggacag tcgtttgccg tctgaatttg acctgagcgc atttttacgc
      781 gccggagaaa accgcctcgc ggtgatggtg ctgcgctgga gtgacggcag ttatctggaa
      841 gatcaggata tgtggcggat gagcggcatt ttccgtgacg tctcgttgct gcataaaccg
      901 actacacaaa tcagcgattt ccatgttgcc actcgcttta atgatgattt cagccgcgct
      961 gtactggagg ctgaagttca gatgtgcggc gagttgcgtg actacctacg ggtaacagtt
     1021 tctttatggc agggtgaaac gcaggtcgcc agcggcaccg cgcctttcgg cggtgaaatt
     1081 atcgatgagc gtggtggtta tgccgatcgc gtcacactac gtctgaacgt cgaaaacccg
     1141 aaactgtgga gcgccgaaat cccgaatctc tatcgtgcgg tggttgaact gcacaccgcc
     1201 gacggcacgc tgattgaagc agaagcctgc gatgtcggtt tccgcgaggt gcggattgaa
     1261 aatggtctgc tgctgctgaa cggcaagccg ttgctgattc gaggcgttaa ccgtcacgag
     1321 catcatcctc tgcatggtca ggtcatggat gagcagacga tggtgcagga tgtaagttta
     1381 aactattcgt tactaactaa ctttaaacat ttaaattttc agatcctgct gatgaagcag
     1441 aacaacttta acgccgtgcg ctgttcgcat tatccgaacc atccgctgtg gtacacgctg
     1501 tgcgaccgct acggcctgta tgtggtggat gaagccaata ttgaaaccca cggcatggtg
     1561 ccaatgaatc gtctgaccga tgatccgcgc tggctaccgg cgatgagcga acgcgtaacg
     1621 cgaatggtgc agcgcgatcg taatcacccg agtgtgatca tctggtcgct ggggaatgaa
     1681 tcaggccacg gcgctaatca cgacgcgctg tatcgctgga tcaaatctgt cgatccttcc
     1741 cgcccggtgc agtatgaagg cggcggagcc gacaccacgg ccaccgatat tatttgcccg
     1801 atgtacgcgc gcgtggatga agaccagccc ttcccggctg tgccgaaatg gtccatcaaa
     1861 aaatggcttt cgctacctgg agagacgcgc ccgctgatcc tttgcgaata cgcccacgcg
     1921 atgggtaaca gtcttggcgg tttcgctaaa tactggcagg cgtttcgtca gtatccccgt
     1981 ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc tgattaaata tgatgaaaac
     2041 ggcaacccgt ggtcggctta cggcggtgat tttggcgata cgccgaacga tcgccagttc
     2101 tgtatgaacg gtctggtctt tgccgaccgc acgccgcatc cagcgtaagt ttaaacaata
     2161 acctactaac taacgtagat aatttaaatt ttcaggctga cggaagcaaa acaccagcag
     2221 cagtttttcc agttccgttt atccgggcaa accatcgaag tgaccagcga atacctgttc
     2281 cgtcatagcg ataacgagct cctgcactgg atggtggcgc tggatggtaa gccgctggca
     2341 agcggtgaag tgcctctgga tgtcgctcca caaggtaaac agttgattga actgcctgaa
     2401 ctaccgcagc cggagagcgc cgggcaactc tggctcacag tacgcgtagt gcaaccgaac
     2461 gcgaccgcat ggtcagaagc cgggcacatc agcgcctggc agcagtggcg tctggcggaa
     2521 aacctcagtg tgacgctccc cgccgcgtcc cacgccatcc cgcatctgac caccagcgaa
     2581 atggattttt gcatcgagct gggtaataag cgttggcaat ttaaccgcca gtcaggcttt
     2641 ctttcacaga tgtggattgg cgataaaaaa caactgctga cgccgctgcg cgatcagttc
     2701 acccgtgcac cgctggataa cgacattggc gtaagtgaag cgacccgcat tgaccctaac
     2761 gcctgggtcg aacgctggaa ggcggcgggc cattaccagg ccgaagcagc gttgttgcag
     2821 tgcacggcag atacacttgc tgatgcggtg ctgattacga ccgctcacgc gtggcagcat
     2881 caggggaaaa ccttatttat cagccggaaa acctaccgga ttgatggtag tggtcaaatg
     2941 gcgattaccg ttgatgttga agtggcgagc gatacaccgc atccggcgcg gattggcctg
     3001 aactgccagc tggcgcaggt agcagagcgg gtaaactggc tcggattagg gccgcaagaa
     3061 aactatcccg accgccttac tgccgcctgt tttgaccgct gggatctgcc attgtcagac
     3121 atgtagtaag tttaaacttg atagtactaa ctaacatgtt tcatttaaat tttcagtacc
     3181 ccgtacgtct tcccgagcga aaacggtctg cgctgcggga cgcgcgaatt gaattatggc
     3241 ccacaccagt ggcgcggcga cttccagttc aacatcagcc gctacagtca acagcaactg
     3301 atggaaacca gccatcgcca tctgctgcac gcggaagaag gcacatggct gaatatcgac
     3361 ggtttccata tggggattgg tggcgacgac tcctggagcc cgtcagtatc ggcggaattc
     3421 caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata ataggggccg
     3481 ctgtcatcag atcgccatct cgcgcccgtg cctctgactt ctaagtccaa ttactcttca
     3541 acatccctac atgctctttc tccctgtgct cccaccccct atttttgtta ttatcaaaaa
     3601 acttctctta atttctttgt tttttagctt cttttaagtc acctctaaca atgaaattgt
     3661 gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt gctccctccc
     3721 cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt
     3781 ttttacttct gataaatttt tttgaaacat catagaaaaa accgcacaca aaatacctta
     3841 tcatatgtta cgtttcagtt tatgaccgca atttttattt cttcgcacgt ctgggcctct
     3901 catgacgtca aatcatgctc atcgtgaaaa agttttggag tatttttgga atttttcaat
     3961 caagtgaaag tttatgaaat taattttcct gcttttgctt tttggggttt cccctattgt
     4021 ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc acaagtattg atgagcacga
     4081 tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga gtagaagttg
     4141 ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag aatacattcc
     4201 caatatacca aacataactg tttcctacta gtcggccgta cgggcccttt cgtctcgcgc
     4261 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     4321 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4381 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     4441 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cggccttaag
     4501 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4561 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4681 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4741 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4801 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4861 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4921 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4981 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5041 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5101 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5161 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5221 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5281 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5341 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5401 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5461 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5521 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5581 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5641 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5701 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5761 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5821 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     5881 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5941 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6001 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6061 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     6121 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6181 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6241 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6301 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6361 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6421 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6481 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6541 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6601 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6661 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagct
//
