LOCUS       sequence-60-expo        7017 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 7017)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7017)
  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: pPD50_14
FEATURES             Location/Qualifiers
     source          1..7017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             576..596
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             684..3731
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(686..708)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4591..4613)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4713..4732
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4810..4828)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4896..5000
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5001..5069,5070..5861)
                     /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(5219..5238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6032..6620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6521..6540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6774..6791
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6908..6929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6944..6949,6950..6967,6968..6974)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6982..6998
                     /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     6987..7009
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7006..7017,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7006..7017,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcaagagcat
       61 ttgaatcaga atatggagaa cggagcatga gcattttcga agttttttag atgcactaga
      121 acaaagcgtg ttggcttcct ctgagcccgc tttccttata tacccgcatt ctgcagcctt
      181 acagaatgtt ctagaaggtc ctagatgcat tcgtttgaaa atactcccgg tgggtgcaaa
      241 gagacgcaga cggaaaatgt atctgggtct ctttattgtg tacactactt ttccatgtac
      301 cgaatgtgag tcgccctcct tttgcaacaa gcagctcgaa tgttctagaa aaaggtggaa
      361 aatagtataa ataccgttga aaataaatac cgaacaacat ttgctctaat tgtgaaatta
      421 gaaatcttca aactataatc atctcactgg atccccggga ttggccaaag gacccaaagg
      481 tatgtttcga atgatactaa cataacatag aacattttca ggaggaccct tggctagagg
      541 atccccgggt accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg
      601 tgggtgaaga ccagaaacag cacctcgaac tgagccgcga tattgcccag cgtttcaacg
      661 cgctgtatgg cgagatcgat cccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
      721 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
      781 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
      841 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
      901 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
      961 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
     1021 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     1081 ttatttttga tggcgttaac tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt
     1141 acggccagga cagtcgtttg ccgtctgaat ttgacctgag cgcattttta cgcgccggag
     1201 aaaaccgcct cgcggtgatg gtgctgcgct ggagtgacgg cagttatctg gaagatcagg
     1261 atatgtggcg gatgagcggc attttccgtg acgtctcgtt gctgcataaa ccgactacac
     1321 aaatcagcga tttccatgtt gccactcgct ttaatgatga tttcagccgc gctgtactgg
     1381 aggctgaagt tcagatgtgc ggcgagttgc gtgactacct acgggtaaca gtttctttat
     1441 ggcagggtga aacgcaggtc gccagcggca ccgcgccttt cggcggtgaa attatcgatg
     1501 agcgtggtgg ttatgccgat cgcgtcacac tacgtctgaa cgtcgaaaac ccgaaactgt
     1561 ggagcgccga aatcccgaat ctctatcgtg cggtggttga actgcacacc gccgacggca
     1621 cgctgattga agcagaagcc tgcgatgtcg gtttccgcga ggtgcggatt gaaaatggtc
     1681 tgctgctgct gaacggcaag ccgttgctga ttcgaggcgt taaccgtcac gagcatcatc
     1741 ctctgcatgg tcaggtcatg gatgagcaga cgatggtgca ggatatcctg ctgatgaagc
     1801 agaacaactt taacgccgtg cgctgttcgc attatccgaa ccatccgctg tggtacacgc
     1861 tgtgcgaccg ctacggcctg tatgtggtgg atgaagccaa tattgaaacc cacggcatgg
     1921 tgccaatgaa tcgtctgacc gatgatccgc gctggctacc ggcgatgagc gaacgcgtaa
     1981 cgcgaatggt gcagcgcgat cgtaatcacc cgagtgtgat catctggtcg ctggggaatg
     2041 aatcaggcca cggcgctaat cacgacgcgc tgtatcgctg gatcaaatct gtcgatcctt
     2101 cccgcccggt gcagtatgaa ggcggcggag ccgacaccac ggccaccgat attatttgcc
     2161 cgatgtacgc gcgcgtggat gaagaccagc ccttcccggc tgtgccgaaa tggtccatca
     2221 aaaaatggct ttcgctacct ggagagacgc gcccgctgat cctttgcgaa tacgcccacg
     2281 cgatgggtaa cagtcttggc ggtttcgcta aatactggca ggcgtttcgt cagtatcccc
     2341 gtttacaggg cggcttcgtc tgggactggg tggatcagtc gctgattaaa tatgatgaaa
     2401 acggcaaccc gtggtcggct tacggcggtg attttggcga tacgccgaac gatcgccagt
     2461 tctgtatgaa cggtctggtc tttgccgacc gcacgccgca tccagcgctg acggaagcaa
     2521 aacaccagca gcagtttttc cagttccgtt tatccgggca aaccatcgaa gtgaccagcg
     2581 aatacctgtt ccgtcatagc gataacgagc tcctgcactg gatggtggcg ctggatggta
     2641 agccgctggc aagcggtgaa gtgcctctgg atgtcgctcc acaaggtaaa cagttgattg
     2701 aactgcctga actaccgcag ccggagagcg ccgggcaact ctggctcaca gtacgcgtag
     2761 tgcaaccgaa cgcgaccgca tggtcagaag ccgggcacat cagcgcctgg cagcagtggc
     2821 gtctggcgga aaacctcagt gtgacgctcc ccgccgcgtc ccacgccatc ccgcatctga
     2881 ccaccagcga aatggatttt tgcatcgagc tgggtaataa gcgttggcaa tttaaccgcc
     2941 agtcaggctt tctttcacag atgtggattg gcgataaaaa acaactgctg acgccgctgc
     3001 gcgatcagtt cacccgtgca ccgctggata acgacattgg cgtaagtgaa gcgacccgca
     3061 ttgaccctaa cgcctgggtc gaacgctgga aggcggcggg ccattaccag gccgaagcag
     3121 cgttgttgca gtgcacggca gatacacttg ctgatgcggt gctgattacg accgctcacg
     3181 cgtggcagca tcaggggaaa accttattta tcagccggaa aacctaccgg attgatggta
     3241 gtggtcaaat ggcgattacc gttgatgttg aagtggcgag cgatacaccg catccggcgc
     3301 ggattggcct gaactgccag ctggcgcagg tagcagagcg ggtaaactgg ctcggattag
     3361 ggccgcaaga aaactatccc gaccgcctta ctgccgcctg ttttgaccgc tgggatctgc
     3421 cattgtcaga catgtatacc ccgtacgtct tcccgagcga aaacggtctg cgctgcggga
     3481 cgcgcgaatt gaattatggc ccacaccagt ggcgcggcga cttccagttc aacatcagcc
     3541 gctacagtca acagcaactg atggaaacca gccatcgcca tctgctgcac gcggaagaag
     3601 gcacatggct gaatatcgac ggtttccata tggggattgg tggcgacgac tcctggagcc
     3661 cgtcagtatc ggcggaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt
     3721 gtcaaaaata ataggcctac tagcgtcgac ggtaccatgg tattgatatc tgagctccgc
     3781 atcggccgct gtcatcagat cgccatctcg cgcccgtgcc tctgacttct aagtccaatt
     3841 actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat ttttgttatt
     3901 atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac ctctaacaat
     3961 gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc
     4021 tccctccccc cattaataat aattctatcc caaaatctac acaatgttct gtgtacactt
     4081 cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac cgcacacaaa
     4141 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct
     4201 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat
     4261 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tggggtttcc
     4321 cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     4381 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     4441 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     4501 tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg
     4561 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4621 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4681 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     4741 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     4801 gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4861 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4921 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4981 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5041 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5101 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5161 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5221 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5281 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5341 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5401 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5461 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5521 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5581 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5641 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5701 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5761 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5821 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5881 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5941 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6001 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6061 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     6181 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6241 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6301 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6361 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6421 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6481 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6541 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6601 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6661 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6721 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6781 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6841 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6901 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6961 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagct
//
