LOCUS       sequence-78-expo        6096 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 6096)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6096)
  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: pPD8_02
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             85..105
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             193..3240
                     /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(195..217)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3344..3363)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3463..3485
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3523..3541)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(4080..4148,4149..4940)
                     /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(4298..4317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5111..5699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5600..5619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5853..5870
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5987..6008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6023..6028,6029..6046,6047..6053)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6061..6077
                     /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     6066..6088
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6085..6096,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6085..6096,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta
       61 ccgagctcag aaaaaatgac tgctccaaag aagaagcgta aggtaccggt gggtgaagac
      121 cagaaacagc acctcgaact gagccgcgat attgcccagc gtttcaacgc gctgtatggc
      181 gagatcgatc ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
      241 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
      301 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg
      361 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc
      421 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc
      481 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg
      541 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat
      601 ggcgttaact cggcgtttca tctgtggtgc aacgggcgct gggtcggtta cggccaggac
      661 agtcgtttgc cgtctgaatt tgacctgagc gcatttttac gcgccggaga aaaccgcctc
      721 gcggtgatgg tgctgcgctg gagtgacggc agttatctgg aagatcagga tatgtggcgg
      781 atgagcggca ttttccgtga cgtctcgttg ctgcataaac cgactacaca aatcagcgat
      841 ttccatgttg ccactcgctt taatgatgat ttcagccgcg ctgtactgga ggctgaagtt
      901 cagatgtgcg gcgagttgcg tgactaccta cgggtaacag tttctttatg gcagggtgaa
      961 acgcaggtcg ccagcggcac cgcgcctttc ggcggtgaaa ttatcgatga gcgtggtggt
     1021 tatgccgatc gcgtcacact acgtctgaac gtcgaaaacc cgaaactgtg gagcgccgaa
     1081 atcccgaatc tctatcgtgc ggtggttgaa ctgcacaccg ccgacggcac gctgattgaa
     1141 gcagaagcct gcgatgtcgg tttccgcgag gtgcggattg aaaatggtct gctgctgctg
     1201 aacggcaagc cgttgctgat tcgaggcgtt aaccgtcacg agcatcatcc tctgcatggt
     1261 caggtcatgg atgagcagac gatggtgcag gatatcctgc tgatgaagca gaacaacttt
     1321 aacgccgtgc gctgttcgca ttatccgaac catccgctgt ggtacacgct gtgcgaccgc
     1381 tacggcctgt atgtggtgga tgaagccaat attgaaaccc acggcatggt gccaatgaat
     1441 cgtctgaccg atgatccgcg ctggctaccg gcgatgagcg aacgcgtaac gcgaatggtg
     1501 cagcgcgatc gtaatcaccc gagtgtgatc atctggtcgc tggggaatga atcaggccac
     1561 ggcgctaatc acgacgcgct gtatcgctgg atcaaatctg tcgatccttc ccgcccggtg
     1621 cagtatgaag gcggcggagc cgacaccacg gccaccgata ttatttgccc gatgtacgcg
     1681 cgcgtggatg aagaccagcc cttcccggct gtgccgaaat ggtccatcaa aaaatggctt
     1741 tcgctacctg gagagacgcg cccgctgatc ctttgcgaat acgcccacgc gatgggtaac
     1801 agtcttggcg gtttcgctaa atactggcag gcgtttcgtc agtatccccg tttacagggc
     1861 ggcttcgtct gggactgggt ggatcagtcg ctgattaaat atgatgaaaa cggcaacccg
     1921 tggtcggctt acggcggtga ttttggcgat acgccgaacg atcgccagtt ctgtatgaac
     1981 ggtctggtct ttgccgaccg cacgccgcat ccagcgctga cggaagcaaa acaccagcag
     2041 cagtttttcc agttccgttt atccgggcaa accatcgaag tgaccagcga atacctgttc
     2101 cgtcatagcg ataacgagct cctgcactgg atggtggcgc tggatggtaa gccgctggca
     2161 agcggtgaag tgcctctgga tgtcgctcca caaggtaaac agttgattga actgcctgaa
     2221 ctaccgcagc cggagagcgc cgggcaactc tggctcacag tacgcgtagt gcaaccgaac
     2281 gcgaccgcat ggtcagaagc cgggcacatc agcgcctggc agcagtggcg tctggcggaa
     2341 aacctcagtg tgacgctccc cgccgcgtcc cacgccatcc cgcatctgac caccagcgaa
     2401 atggattttt gcatcgagct gggtaataag cgttggcaat ttaaccgcca gtcaggcttt
     2461 ctttcacaga tgtggattgg cgataaaaaa caactgctga cgccgctgcg cgatcagttc
     2521 acccgtgcac cgctggataa cgacattggc gtaagtgaag cgacccgcat tgaccctaac
     2581 gcctgggtcg aacgctggaa ggcggcgggc cattaccagg ccgaagcagc gttgttgcag
     2641 tgcacggcag atacacttgc tgatgcggtg ctgattacga ccgctcacgc gtggcagcat
     2701 caggggaaaa ccttatttat cagccggaaa acctaccgga ttgatggtag tggtcaaatg
     2761 gcgattaccg ttgatgttga agtggcgagc gatacaccgc atccggcgcg gattggcctg
     2821 aactgccagc tggcgcaggt agcagagcgg gtaaactggc tcggattagg gccgcaagaa
     2881 aactatcccg accgccttac tgccgcctgt tttgaccgct gggatctgcc attgtcagac
     2941 atgtataccc cgtacgtctt cccgagcgaa aacggtctgc gctgcgggac gcgcgaattg
     3001 aattatggcc cacaccagtg gcgcggcgac ttccagttca acatcagccg ctacagtcaa
     3061 cagcaactga tggaaaccag ccatcgccat ctgctgcacg cggaagaagg cacatggctg
     3121 aatatcgacg gtttccatat ggggattggt ggcgacgact cctggagccc gtcagtatcg
     3181 gcggaattcc aactgagcgc cggtcgctac cattaccaac ttgtctggtg tcaaaaataa
     3241 taggcctact agtcggccgt acgggccctt aaggccgcct gatgcggtat tttctcctta
     3301 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     3361 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3421 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3481 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3541 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3661 tcatgagaca ataaccctga taaatgcttc aataatacaa ttttcagaat acgttttttg
     3721 tgggcttggg tatattgttt ttaatgttat acttgcagtc gtgaaatttg attttcaaat
     3781 ttgtagaaaa atcaagaaaa taattgcaac attcgcttgt gtcaaaaacc aatttcaaca
     3841 aattttcgtg tgagaaatac attaccagaa ggcatttttt cacacgatta gcattttgga
     3901 ctactttatt aaatttttgc gtgtaatttt gaattaaatt gtattatatt actacttaaa
     3961 aaacaaaaaa tttgaccact gagcggatcg aacgcccaac ctttcgatct agagtcgaac
     4021 gcgctaccat tgcgccaagc agtcatgtta ttctctcttg tcattgaaaa aggaagagta
     4081 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4141 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4201 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4261 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4321 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4381 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4441 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4501 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4561 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4621 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4681 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4741 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4801 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4861 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4921 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4981 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5041 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5101 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5161 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5221 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5281 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5341 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5401 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5461 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5521 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5581 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5641 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5701 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5761 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5821 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5881 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5941 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     6001 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     6061 ttgtgagcgg ataacaattt cacacaggaa acagct
//
