LOCUS       sequence-45-expo        7003 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 7003)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7003)
  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: pPD16_51
FEATURES             Location/Qualifiers
     source          1..7003
                     /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(4577..4599)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4699..4718
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4796..4814)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4882..4986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4987..5055,5056..5847)
                     /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(5205..5224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6018..6606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6507..6526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6760..6777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6894..6915
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6930..6935,6936..6953,6954..6960)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6968..6984
                     /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     6973..6995
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6992..7003,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6992..7003,1..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 taggggccgc tgtcatcaga tcgccatctc gcgcccgtgc ctctgacttc taagtccaat
     3301 tactcttcaa catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat
     3361 tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc ttttaagtca cctctaacaa
     3421 tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg
     3481 ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact
     3541 tcttatgttt tttacttctg ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa
     3601 aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc
     3661 tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa
     3721 tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttggggtttc
     3781 ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca caagtattga
     3841 tgagcacgat gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag
     3901 tagaagttga taatttgaaa gtggagtagt gtctatgggg tttttgcctt aaatgacaga
     3961 atacattccc aatataccaa acataactgt ttaaaattaa acatttttct aaattttata
     4021 tgatttcttt taaatttgca aaaattactt aaatttgaat tcccgcgcaa atgagtgact
     4081 tcattttctg cattattgtg ttttccggct atattaatag gtatttgttt gtgtttttct
     4141 ttattttatg attcgaactc caatttgtaa attttcgaac atatttccct aaagaaaaaa
     4201 tatgattaat ctggaaaaat tggaaaatta tttttcaaat aaaaaacaaa gaaaaaaatg
     4261 aagaaaaacc tattagtttg gccataaaac gcaaaaatgt cgaaaatgac gtcactcatc
     4321 tgcgcgggaa atcaagaata attcggcctt ttttattttt ttggaaaatc gtaaaacatt
     4381 tagaaaaatt ttttaatagt tatagtggga ctgtattctg tcatttaggg caaaagccag
     4441 agacgctact ccaccgttaa catgaattat gaatattatt gcgacaagac ccaaacattg
     4501 ataaaccgca aatctagcct actagtcggc cgtacgggcc ctttcgtctc gcgcgtttcg
     4561 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     4621 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     4681 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     4741 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcggcct taagggcctc
     4801 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4861 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4921 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4981 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5041 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5101 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5161 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5221 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5281 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5341 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5401 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5461 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5521 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5581 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5641 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5701 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5761 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5821 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5881 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5941 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6001 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6061 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     6181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     6421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6781 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6841 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     6901 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     6961 tgtggaattg tgagcggata acaatttcac acaggaaaca gct
//
