LOCUS       sequence-80-expo        6603 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 6603)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6603)
  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_23
FEATURES             Location/Qualifiers
     source          1..6603
                     /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"
     CDS             271..3318
                     /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(273..295)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4177..4199)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4299..4318
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4396..4414)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4482..4586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4587..4655,4656..5447)
                     /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(4805..4824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5618..6206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6107..6126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6360..6377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6494..6515
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6530..6535,6536..6553,6554..6560)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6568..6584
                     /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     6573..6595
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6592..6603,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6592..6603,1..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 tatcctgctg
     1381 atgaagcaga acaactttaa cgccgtgcgc tgttcgcatt atccgaacca tccgctgtgg
     1441 tacacgctgt gcgaccgcta cggcctgtat gtggtggatg aagccaatat tgaaacccac
     1501 ggcatggtgc caatgaatcg tctgaccgat gatccgcgct ggctaccggc gatgagcgaa
     1561 cgcgtaacgc gaatggtgca gcgcgatcgt aatcacccga gtgtgatcat ctggtcgctg
     1621 gggaatgaat caggccacgg cgctaatcac gacgcgctgt atcgctggat caaatctgtc
     1681 gatccttccc gcccggtgca gtatgaaggc ggcggagccg acaccacggc caccgatatt
     1741 atttgcccga tgtacgcgcg cgtggatgaa gaccagccct tcccggctgt gccgaaatgg
     1801 tccatcaaaa aatggctttc gctacctgga gagacgcgcc cgctgatcct ttgcgaatac
     1861 gcccacgcga tgggtaacag tcttggcggt ttcgctaaat actggcaggc gtttcgtcag
     1921 tatccccgtt tacagggcgg cttcgtctgg gactgggtgg atcagtcgct gattaaatat
     1981 gatgaaaacg gcaacccgtg gtcggcttac ggcggtgatt ttggcgatac gccgaacgat
     2041 cgccagttct gtatgaacgg tctggtcttt gccgaccgca cgccgcatcc agcgctgacg
     2101 gaagcaaaac accagcagca gtttttccag ttccgtttat ccgggcaaac catcgaagtg
     2161 accagcgaat acctgttccg tcatagcgat aacgagctcc tgcactggat ggtggcgctg
     2221 gatggtaagc cgctggcaag cggtgaagtg cctctggatg tcgctccaca aggtaaacag
     2281 ttgattgaac tgcctgaact accgcagccg gagagcgccg ggcaactctg gctcacagta
     2341 cgcgtagtgc aaccgaacgc gaccgcatgg tcagaagccg ggcacatcag cgcctggcag
     2401 cagtggcgtc tggcggaaaa cctcagtgtg acgctccccg ccgcgtccca cgccatcccg
     2461 catctgacca ccagcgaaat ggatttttgc atcgagctgg gtaataagcg ttggcaattt
     2521 aaccgccagt caggctttct ttcacagatg tggattggcg ataaaaaaca actgctgacg
     2581 ccgctgcgcg atcagttcac ccgtgcaccg ctggataacg acattggcgt aagtgaagcg
     2641 acccgcattg accctaacgc ctgggtcgaa cgctggaagg cggcgggcca ttaccaggcc
     2701 gaagcagcgt tgttgcagtg cacggcagat acacttgctg atgcggtgct gattacgacc
     2761 gctcacgcgt ggcagcatca ggggaaaacc ttatttatca gccggaaaac ctaccggatt
     2821 gatggtagtg gtcaaatggc gattaccgtt gatgttgaag tggcgagcga tacaccgcat
     2881 ccggcgcgga ttggcctgaa ctgccagctg gcgcaggtag cagagcgggt aaactggctc
     2941 ggattagggc cgcaagaaaa ctatcccgac cgccttactg ccgcctgttt tgaccgctgg
     3001 gatctgccat tgtcagacat gtataccccg tacgtcttcc cgagcgaaaa cggtctgcgc
     3061 tgcgggacgc gcgaattgaa ttatggccca caccagtggc gcggcgactt ccagttcaac
     3121 atcagccgct acagtcaaca gcaactgatg gaaaccagcc atcgccatct gctgcacgcg
     3181 gaagaaggca catggctgaa tatcgacggt ttccatatgg ggattggtgg cgacgactcc
     3241 tggagcccgt cagtatcggc ggaattccaa ctgagcgccg gtcgctacca ttaccaactt
     3301 gtctggtgtc aaaaataata ggggccgctg tcatcagagt aagtttaaac tgagttctac
     3361 taactaacga gtaatattta aattttcagc atctcgcgcc cgtgcctctg acttctaagt
     3421 ccaattactc ttcaacatcc ctacatgctc tttctccctg tgctcccacc ccctattttt
     3481 gttattatca aaaaacttct cttaatttct ttgtttttta gcttctttta agtcacctct
     3541 aacaatgaaa ttgtgtagat tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa
     3601 ttgtgctccc tccccccatt aataataatt ctatcccaaa atctacacaa tgttctgtgt
     3661 acacttctta tgttttttac ttctgataaa tttttttgaa acatcataga aaaaaccgca
     3721 cacaaaatac cttatcatat gttacgtttc agtttatgac cgcaattttt atttcttcgc
     3781 acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt
     3841 tggaattttt caatcaagtg aaagtttatg aaattaattt tcctgctttt gctttttggg
     3901 gtttccccta ttgtttgtca agatttcgag gacggcgttt ttcttgctaa aatcacaagt
     3961 attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc tcagtggaag
     4021 gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt gccttaaatg
     4081 acagaataca ttcccaatat accaaacata actgtttcct actagtcggc cgtacgggcc
     4141 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4201 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4261 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     4321 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     4381 caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4441 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4501 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4561 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4621 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4681 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4741 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4801 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4861 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4921 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4981 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5041 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5101 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5161 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5221 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5281 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5341 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5401 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5461 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5521 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5581 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5641 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5701 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5761 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5821 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5881 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5941 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6001 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6061 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6121 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6181 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6241 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6301 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6361 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     6421 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     6481 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     6541 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6601 gct
//
