LOCUS       sequence-69-expo        6779 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 6779)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6779)
  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: pPD87_18
FEATURES             Location/Qualifiers
     source          1..6779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             386..406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             494..3541
                     /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(496..518)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4353..4375)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4475..4494
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4572..4590)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4658..4762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4763..4831,4832..5623)
                     /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(4981..5000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5794..6382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6283..6302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6536..6553
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6670..6691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6706..6711,6712..6729,6730..6736)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6744..6760
                     /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     6749..6771
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6768..6779,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6768..6779,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccaattc
       61 ttgagacttc cactatcttc tctggcaccc tccactacgc gtaggcgtct ctcgcttcgt
      121 gtattcccgg gaagccgggt tcccgtctct cccgccgctg ccgctgccgc acacagcttt
      181 acacctcgta gaatccccaa agaggggcgt ggcttgcggg tgccaacatc ctcctgccga
      241 ggaagaagca ggcactcatc actcgcatca tcaacctcgg gattggccaa aggacccaaa
      301 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      361 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tgggtgaaga
      421 ccagaaacag cacctcgaac tgagccgcga tattgcccag cgtttcaacg cgctgtatgg
      481 cgagatcgat cccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      541 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      601 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc
      661 ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt
      721 cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac
      781 ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc
      841 gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga
      901 tggcgttaac tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt acggccagga
      961 cagtcgtttg ccgtctgaat ttgacctgag cgcattttta cgcgccggag aaaaccgcct
     1021 cgcggtgatg gtgctgcgct ggagtgacgg cagttatctg gaagatcagg atatgtggcg
     1081 gatgagcggc attttccgtg acgtctcgtt gctgcataaa ccgactacac aaatcagcga
     1141 tttccatgtt gccactcgct ttaatgatga tttcagccgc gctgtactgg aggctgaagt
     1201 tcagatgtgc ggcgagttgc gtgactacct acgggtaaca gtttctttat ggcagggtga
     1261 aacgcaggtc gccagcggca ccgcgccttt cggcggtgaa attatcgatg agcgtggtgg
     1321 ttatgccgat cgcgtcacac tacgtctgaa cgtcgaaaac ccgaaactgt ggagcgccga
     1381 aatcccgaat ctctatcgtg cggtggttga actgcacacc gccgacggca cgctgattga
     1441 agcagaagcc tgcgatgtcg gtttccgcga ggtgcggatt gaaaatggtc tgctgctgct
     1501 gaacggcaag ccgttgctga ttcgaggcgt taaccgtcac gagcatcatc ctctgcatgg
     1561 tcaggtcatg gatgagcaga cgatggtgca ggatatcctg ctgatgaagc agaacaactt
     1621 taacgccgtg cgctgttcgc attatccgaa ccatccgctg tggtacacgc tgtgcgaccg
     1681 ctacggcctg tatgtggtgg atgaagccaa tattgaaacc cacggcatgg tgccaatgaa
     1741 tcgtctgacc gatgatccgc gctggctacc ggcgatgagc gaacgcgtaa cgcgaatggt
     1801 gcagcgcgat cgtaatcacc cgagtgtgat catctggtcg ctggggaatg aatcaggcca
     1861 cggcgctaat cacgacgcgc tgtatcgctg gatcaaatct gtcgatcctt cccgcccggt
     1921 gcagtatgaa ggcggcggag ccgacaccac ggccaccgat attatttgcc cgatgtacgc
     1981 gcgcgtggat gaagaccagc ccttcccggc tgtgccgaaa tggtccatca aaaaatggct
     2041 ttcgctacct ggagagacgc gcccgctgat cctttgcgaa tacgcccacg cgatgggtaa
     2101 cagtcttggc ggtttcgcta aatactggca ggcgtttcgt cagtatcccc gtttacaggg
     2161 cggcttcgtc tgggactggg tggatcagtc gctgattaaa tatgatgaaa acggcaaccc
     2221 gtggtcggct tacggcggtg attttggcga tacgccgaac gatcgccagt tctgtatgaa
     2281 cggtctggtc tttgccgacc gcacgccgca tccagcgctg acggaagcaa aacaccagca
     2341 gcagtttttc cagttccgtt tatccgggca aaccatcgaa gtgaccagcg aatacctgtt
     2401 ccgtcatagc gataacgagc tcctgcactg gatggtggcg ctggatggta agccgctggc
     2461 aagcggtgaa gtgcctctgg atgtcgctcc acaaggtaaa cagttgattg aactgcctga
     2521 actaccgcag ccggagagcg ccgggcaact ctggctcaca gtacgcgtag tgcaaccgaa
     2581 cgcgaccgca tggtcagaag ccgggcacat cagcgcctgg cagcagtggc gtctggcgga
     2641 aaacctcagt gtgacgctcc ccgccgcgtc ccacgccatc ccgcatctga ccaccagcga
     2701 aatggatttt tgcatcgagc tgggtaataa gcgttggcaa tttaaccgcc agtcaggctt
     2761 tctttcacag atgtggattg gcgataaaaa acaactgctg acgccgctgc gcgatcagtt
     2821 cacccgtgca ccgctggata acgacattgg cgtaagtgaa gcgacccgca ttgaccctaa
     2881 cgcctgggtc gaacgctgga aggcggcggg ccattaccag gccgaagcag cgttgttgca
     2941 gtgcacggca gatacacttg ctgatgcggt gctgattacg accgctcacg cgtggcagca
     3001 tcaggggaaa accttattta tcagccggaa aacctaccgg attgatggta gtggtcaaat
     3061 ggcgattacc gttgatgttg aagtggcgag cgatacaccg catccggcgc ggattggcct
     3121 gaactgccag ctggcgcagg tagcagagcg ggtaaactgg ctcggattag ggccgcaaga
     3181 aaactatccc gaccgcctta ctgccgcctg ttttgaccgc tgggatctgc cattgtcaga
     3241 catgtatacc ccgtacgtct tcccgagcga aaacggtctg cgctgcggga cgcgcgaatt
     3301 gaattatggc ccacaccagt ggcgcggcga cttccagttc aacatcagcc gctacagtca
     3361 acagcaactg atggaaacca gccatcgcca tctgctgcac gcggaagaag gcacatggct
     3421 gaatatcgac ggtttccata tggggattgg tggcgacgac tcctggagcc cgtcagtatc
     3481 ggcggaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata
     3541 ataggggccg ctgtcatcag atcgccatct cgcgcccgtg cctctgactt ctaagtccaa
     3601 ttactcttca acatccctac atgctctttc tccctgtgct cccaccccct atttttgtta
     3661 ttatcaaaaa acttctctta atttctttgt tttttagctt cttttaagtc acctctaaca
     3721 atgaaattgt gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt
     3781 gctccctccc cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac
     3841 ttcttatgtt ttttacttct gataaatttt tttgaaacat catagaaaaa accgcacaca
     3901 aaatacctta tcatatgtta cgtttcagtt tatgaccgca atttttattt cttcgcacgt
     3961 ctgggcctct catgacgtca aatcatgctc atcgtgaaaa agttttggag tatttttgga
     4021 atttttcaat caagtgaaag tttatgaaat taattttcct gcttttgctt tttggggttt
     4081 cccctattgt ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc acaagtattg
     4141 atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga
     4201 gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag
     4261 aatacattcc caatatacca aacataactg tttcctacta gtcggccgta cgggcccttt
     4321 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4381 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4441 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4501 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     4561 cggccttaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4621 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4681 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4741 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4801 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4861 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4921 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4981 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5041 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5101 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5161 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5221 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5281 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5341 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5401 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5461 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5521 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5581 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5641 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5701 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5941 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6181 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6421 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6481 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6541 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6601 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     6661 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     6721 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagct
//
