LOCUS       sequence-50-expo        6555 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 6555)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6555)
  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: pPD22_04
FEATURES             Location/Qualifiers
     source          1..6555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             162..182
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             270..3317
                     /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(272..294)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4129..4151)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4251..4270
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4348..4366)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4434..4538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4539..4607,4608..5399)
                     /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(4757..4776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5570..6158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6059..6078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6312..6329
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6446..6467
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6482..6487,6488..6505,6506..6512)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6520..6536
                     /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     6525..6547
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6544..6555,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6544..6555,1..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 gagggtaccg agctcagaaa aaatgactgc tccaaagaag aagcgtaagg
      181 taccggtggg tgaagaccag aaacagcacc tcgaactgag ccgcgatatt gcccagcgtt
      241 tcaacgcgct gtatggcgag atcgatcccg tcgttttaca acgtcgtgac tgggaaaacc
      301 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      361 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      421 gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc
      481 ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca
      541 tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc
      601 cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga
      661 cgcgaattat ttttgatggc gttaactcgg cgtttcatct gtggtgcaac gggcgctggg
      721 tcggttacgg ccaggacagt cgtttgccgt ctgaatttga cctgagcgca tttttacgcg
      781 ccggagaaaa ccgcctcgcg gtgatggtgc tgcgctggag tgacggcagt tatctggaag
      841 atcaggatat gtggcggatg agcggcattt tccgtgacgt ctcgttgctg cataaaccga
      901 ctacacaaat cagcgatttc catgttgcca ctcgctttaa tgatgatttc agccgcgctg
      961 tactggaggc tgaagttcag atgtgcggcg agttgcgtga ctacctacgg gtaacagttt
     1021 ctttatggca gggtgaaacg caggtcgcca gcggcaccgc gcctttcggc ggtgaaatta
     1081 tcgatgagcg tggtggttat gccgatcgcg tcacactacg tctgaacgtc gaaaacccga
     1141 aactgtggag cgccgaaatc ccgaatctct atcgtgcggt ggttgaactg cacaccgccg
     1201 acggcacgct gattgaagca gaagcctgcg atgtcggttt ccgcgaggtg cggattgaaa
     1261 atggtctgct gctgctgaac ggcaagccgt tgctgattcg aggcgttaac cgtcacgagc
     1321 atcatcctct gcatggtcag gtcatggatg agcagacgat ggtgcaggat atcctgctga
     1381 tgaagcagaa caactttaac gccgtgcgct gttcgcatta tccgaaccat ccgctgtggt
     1441 acacgctgtg cgaccgctac ggcctgtatg tggtggatga agccaatatt gaaacccacg
     1501 gcatggtgcc aatgaatcgt ctgaccgatg atccgcgctg gctaccggcg atgagcgaac
     1561 gcgtaacgcg aatggtgcag cgcgatcgta atcacccgag tgtgatcatc tggtcgctgg
     1621 ggaatgaatc aggccacggc gctaatcacg acgcgctgta tcgctggatc aaatctgtcg
     1681 atccttcccg cccggtgcag tatgaaggcg gcggagccga caccacggcc accgatatta
     1741 tttgcccgat gtacgcgcgc gtggatgaag accagccctt cccggctgtg ccgaaatggt
     1801 ccatcaaaaa atggctttcg ctacctggag agacgcgccc gctgatcctt tgcgaatacg
     1861 cccacgcgat gggtaacagt cttggcggtt tcgctaaata ctggcaggcg tttcgtcagt
     1921 atccccgttt acagggcggc ttcgtctggg actgggtgga tcagtcgctg attaaatatg
     1981 atgaaaacgg caacccgtgg tcggcttacg gcggtgattt tggcgatacg ccgaacgatc
     2041 gccagttctg tatgaacggt ctggtctttg ccgaccgcac gccgcatcca gcgctgacgg
     2101 aagcaaaaca ccagcagcag tttttccagt tccgtttatc cgggcaaacc atcgaagtga
     2161 ccagcgaata cctgttccgt catagcgata acgagctcct gcactggatg gtggcgctgg
     2221 atggtaagcc gctggcaagc ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt
     2281 tgattgaact gcctgaacta ccgcagccgg agagcgccgg gcaactctgg ctcacagtac
     2341 gcgtagtgca accgaacgcg accgcatggt cagaagccgg gcacatcagc gcctggcagc
     2401 agtggcgtct ggcggaaaac ctcagtgtga cgctccccgc cgcgtcccac gccatcccgc
     2461 atctgaccac cagcgaaatg gatttttgca tcgagctggg taataagcgt tggcaattta
     2521 accgccagtc aggctttctt tcacagatgt ggattggcga taaaaaacaa ctgctgacgc
     2581 cgctgcgcga tcagttcacc cgtgcaccgc tggataacga cattggcgta agtgaagcga
     2641 cccgcattga ccctaacgcc tgggtcgaac gctggaaggc ggcgggccat taccaggccg
     2701 aagcagcgtt gttgcagtgc acggcagata cacttgctga tgcggtgctg attacgaccg
     2761 ctcacgcgtg gcagcatcag gggaaaacct tatttatcag ccggaaaacc taccggattg
     2821 atggtagtgg tcaaatggcg attaccgttg atgttgaagt ggcgagcgat acaccgcatc
     2881 cggcgcggat tggcctgaac tgccagctgg cgcaggtagc agagcgggta aactggctcg
     2941 gattagggcc gcaagaaaac tatcccgacc gccttactgc cgcctgtttt gaccgctggg
     3001 atctgccatt gtcagacatg tataccccgt acgtcttccc gagcgaaaac ggtctgcgct
     3061 gcgggacgcg cgaattgaat tatggcccac accagtggcg cggcgacttc cagttcaaca
     3121 tcagccgcta cagtcaacag caactgatgg aaaccagcca tcgccatctg ctgcacgcgg
     3181 aagaaggcac atggctgaat atcgacggtt tccatatggg gattggtggc gacgactcct
     3241 ggagcccgtc agtatcggcg gaattccaac tgagcgccgg tcgctaccat taccaacttg
     3301 tctggtgtca aaaataatag gggccgctgt catcagatcg ccatctcgcg cccgtgcctc
     3361 tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca
     3421 ccccctattt ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt
     3481 taagtcacct ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa
     3541 agtcgaaaaa aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac
     3601 aatgttctgt gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata
     3661 gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt
     3721 ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt
     3781 ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt
     3841 ttgctttttg gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct
     3901 aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag
     3961 gctcagtgga aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt
     4021 ttgccttaaa tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg
     4081 gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4141 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4201 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4261 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     4321 aaaataccgc atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg
     4381 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4501 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4561 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4621 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4681 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4741 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4801 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4861 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4921 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4981 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5041 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5101 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5161 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5221 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5281 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5341 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5401 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5461 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5521 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5701 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5761 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5821 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5881 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5941 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6001 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6061 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6121 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6181 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6241 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6301 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     6361 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     6421 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     6481 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     6541 acacaggaaa cagct
//
