LOCUS       sequence-51-expo        6560 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 6560)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6560)
  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_11
FEATURES             Location/Qualifiers
     source          1..6560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             167..187
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             275..3322
                     /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(277..299)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4134..4156)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4256..4275
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4353..4371)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4439..4543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4544..4612,4613..5404)
                     /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(4762..4781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5575..6163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6064..6083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6317..6334
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6451..6472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6487..6492,6493..6510,6511..6517)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6525..6541
                     /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     6530..6552
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6549..6560,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6549..6560,1..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 gcttggaggg taccgagctc agaaaaaatg actgctccaa agaagaagcg
      181 taaggtaccg gtgggtgaag accagaaaca gcacctcgaa ctgagccgcg atattgccca
      241 gcgtttcaac gcgctgtatg gcgagatcga tcccgtcgtt ttacaacgtc gtgactggga
      301 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
      361 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
      421 atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga
      481 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
      541 gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga
      601 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
      661 ccagacgcga attatttttg atggcgttaa ctcggcgttt catctgtggt gcaacgggcg
      721 ctgggtcggt tacggccagg acagtcgttt gccgtctgaa tttgacctga gcgcattttt
      781 acgcgccgga gaaaaccgcc tcgcggtgat ggtgctgcgc tggagtgacg gcagttatct
      841 ggaagatcag gatatgtggc ggatgagcgg cattttccgt gacgtctcgt tgctgcataa
      901 accgactaca caaatcagcg atttccatgt tgccactcgc tttaatgatg atttcagccg
      961 cgctgtactg gaggctgaag ttcagatgtg cggcgagttg cgtgactacc tacgggtaac
     1021 agtttcttta tggcagggtg aaacgcaggt cgccagcggc accgcgcctt tcggcggtga
     1081 aattatcgat gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa
     1141 cccgaaactg tggagcgccg aaatcccgaa tctctatcgt gcggtggttg aactgcacac
     1201 cgccgacggc acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat
     1261 tgaaaatggt ctgctgctgc tgaacggcaa gccgttgctg attcgaggcg ttaaccgtca
     1321 cgagcatcat cctctgcatg gtcaggtcat ggatgagcag acgatggtgc aggatatcct
     1381 gctgatgaag cagaacaact ttaacgccgt gcgctgttcg cattatccga accatccgct
     1441 gtggtacacg ctgtgcgacc gctacggcct gtatgtggtg gatgaagcca atattgaaac
     1501 ccacggcatg gtgccaatga atcgtctgac cgatgatccg cgctggctac cggcgatgag
     1561 cgaacgcgta acgcgaatgg tgcagcgcga tcgtaatcac ccgagtgtga tcatctggtc
     1621 gctggggaat gaatcaggcc acggcgctaa tcacgacgcg ctgtatcgct ggatcaaatc
     1681 tgtcgatcct tcccgcccgg tgcagtatga aggcggcgga gccgacacca cggccaccga
     1741 tattatttgc ccgatgtacg cgcgcgtgga tgaagaccag cccttcccgg ctgtgccgaa
     1801 atggtccatc aaaaaatggc tttcgctacc tggagagacg cgcccgctga tcctttgcga
     1861 atacgcccac gcgatgggta acagtcttgg cggtttcgct aaatactggc aggcgtttcg
     1921 tcagtatccc cgtttacagg gcggcttcgt ctgggactgg gtggatcagt cgctgattaa
     1981 atatgatgaa aacggcaacc cgtggtcggc ttacggcggt gattttggcg atacgccgaa
     2041 cgatcgccag ttctgtatga acggtctggt ctttgccgac cgcacgccgc atccagcgct
     2101 gacggaagca aaacaccagc agcagttttt ccagttccgt ttatccgggc aaaccatcga
     2161 agtgaccagc gaatacctgt tccgtcatag cgataacgag ctcctgcact ggatggtggc
     2221 gctggatggt aagccgctgg caagcggtga agtgcctctg gatgtcgctc cacaaggtaa
     2281 acagttgatt gaactgcctg aactaccgca gccggagagc gccgggcaac tctggctcac
     2341 agtacgcgta gtgcaaccga acgcgaccgc atggtcagaa gccgggcaca tcagcgcctg
     2401 gcagcagtgg cgtctggcgg aaaacctcag tgtgacgctc cccgccgcgt cccacgccat
     2461 cccgcatctg accaccagcg aaatggattt ttgcatcgag ctgggtaata agcgttggca
     2521 atttaaccgc cagtcaggct ttctttcaca gatgtggatt ggcgataaaa aacaactgct
     2581 gacgccgctg cgcgatcagt tcacccgtgc accgctggat aacgacattg gcgtaagtga
     2641 agcgacccgc attgacccta acgcctgggt cgaacgctgg aaggcggcgg gccattacca
     2701 ggccgaagca gcgttgttgc agtgcacggc agatacactt gctgatgcgg tgctgattac
     2761 gaccgctcac gcgtggcagc atcaggggaa aaccttattt atcagccgga aaacctaccg
     2821 gattgatggt agtggtcaaa tggcgattac cgttgatgtt gaagtggcga gcgatacacc
     2881 gcatccggcg cggattggcc tgaactgcca gctggcgcag gtagcagagc gggtaaactg
     2941 gctcggatta gggccgcaag aaaactatcc cgaccgcctt actgccgcct gttttgaccg
     3001 ctgggatctg ccattgtcag acatgtatac cccgtacgtc ttcccgagcg aaaacggtct
     3061 gcgctgcggg acgcgcgaat tgaattatgg cccacaccag tggcgcggcg acttccagtt
     3121 caacatcagc cgctacagtc aacagcaact gatggaaacc agccatcgcc atctgctgca
     3181 cgcggaagaa ggcacatggc tgaatatcga cggtttccat atggggattg gtggcgacga
     3241 ctcctggagc ccgtcagtat cggcggaatt ccaactgagc gccggtcgct accattacca
     3301 acttgtctgg tgtcaaaaat aataggggcc gctgtcatca gatcgccatc tcgcgcccgt
     3361 gcctctgact tctaagtcca attactcttc aacatcccta catgctcttt ctccctgtgc
     3421 tcccaccccc tatttttgtt attatcaaaa aacttctctt aatttctttg ttttttagct
     3481 tcttttaagt cacctctaac aatgaaattg tgtagattca aaaatagaat taattcgtaa
     3541 taaaaagtcg aaaaaaattg tgctccctcc ccccattaat aataattcta tcccaaaatc
     3601 tacacaatgt tctgtgtaca cttcttatgt tttttacttc tgataaattt ttttgaaaca
     3661 tcatagaaaa aaccgcacac aaaatacctt atcatatgtt acgtttcagt ttatgaccgc
     3721 aatttttatt tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa
     3781 aagttttgga gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc
     3841 tgcttttgct ttttggggtt tcccctattg tttgtcaaga tttcgaggac ggcgtttttc
     3901 ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa gatcggaaga aggtttgggt
     3961 ttgaggctca gtggaaggtg agtagaagtt gataatttga aagtggagta gtgtctatgg
     4021 ggtttttgcc ttaaatgaca gaatacattc ccaatatacc aaacataact gtttcctact
     4081 agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4141 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4201 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     4261 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     4321 aggagaaaat accgcatcag gcggccttaa gggcctcgtg atacgcctat ttttataggt
     4381 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4441 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4501 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4561 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4621 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4681 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4741 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4801 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4861 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4921 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4981 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5041 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5101 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5161 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5221 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5281 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5341 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5401 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5461 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5521 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5641 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5701 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5941 cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa
     6001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6241 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6301 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6361 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6421 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6481 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6541 atttcacaca ggaaacagct
//
