LOCUS       sequence-71-expo        6826 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 6826)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6826)
  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: pPD88_22
FEATURES             Location/Qualifiers
     source          1..6826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             386..406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             494..3541
                     /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(496..518)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4400..4422)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4522..4541
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4619..4637)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4705..4809
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4810..4878,4879..5670)
                     /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(5028..5047)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5841..6429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6330..6349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6583..6600
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6717..6738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6753..6758,6759..6776,6777..6783)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6791..6807
                     /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     6796..6818
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6815..6826,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6815..6826,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccaattc
       61 ttgagacttc cactatcttc tctggcaccc tccactacgc gtaggcgtct ctcgcttcgt
      121 gtattcccgg gaagccgggt tcccgtctct cccgccgctg ccgctgccgc acacagcttt
      181 acacctcgta gaatccccaa agaggggcgt ggcttgcggg tgccaacatc ctcctgccga
      241 ggaagaagca ggcactcatc actcgcatca tcaacctcgg gattggccaa aggacccaaa
      301 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      361 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tgggtgaaga
      421 ccagaaacag cacctcgaac tgagccgcga tattgcccag cgtttcaacg cgctgtatgg
      481 cgagatcgat cccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      541 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      601 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc
      661 ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt
      721 cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac
      781 ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc
      841 gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga
      901 tggcgttaac tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt acggccagga
      961 cagtcgtttg ccgtctgaat ttgacctgag cgcattttta cgcgccggag aaaaccgcct
     1021 cgcggtgatg gtgctgcgct ggagtgacgg cagttatctg gaagatcagg atatgtggcg
     1081 gatgagcggc attttccgtg acgtctcgtt gctgcataaa ccgactacac aaatcagcga
     1141 tttccatgtt gccactcgct ttaatgatga tttcagccgc gctgtactgg aggctgaagt
     1201 tcagatgtgc ggcgagttgc gtgactacct acgggtaaca gtttctttat ggcagggtga
     1261 aacgcaggtc gccagcggca ccgcgccttt cggcggtgaa attatcgatg agcgtggtgg
     1321 ttatgccgat cgcgtcacac tacgtctgaa cgtcgaaaac ccgaaactgt ggagcgccga
     1381 aatcccgaat ctctatcgtg cggtggttga actgcacacc gccgacggca cgctgattga
     1441 agcagaagcc tgcgatgtcg gtttccgcga ggtgcggatt gaaaatggtc tgctgctgct
     1501 gaacggcaag ccgttgctga ttcgaggcgt taaccgtcac gagcatcatc ctctgcatgg
     1561 tcaggtcatg gatgagcaga cgatggtgca ggatatcctg ctgatgaagc agaacaactt
     1621 taacgccgtg cgctgttcgc attatccgaa ccatccgctg tggtacacgc tgtgcgaccg
     1681 ctacggcctg tatgtggtgg atgaagccaa tattgaaacc cacggcatgg tgccaatgaa
     1741 tcgtctgacc gatgatccgc gctggctacc ggcgatgagc gaacgcgtaa cgcgaatggt
     1801 gcagcgcgat cgtaatcacc cgagtgtgat catctggtcg ctggggaatg aatcaggcca
     1861 cggcgctaat cacgacgcgc tgtatcgctg gatcaaatct gtcgatcctt cccgcccggt
     1921 gcagtatgaa ggcggcggag ccgacaccac ggccaccgat attatttgcc cgatgtacgc
     1981 gcgcgtggat gaagaccagc ccttcccggc tgtgccgaaa tggtccatca aaaaatggct
     2041 ttcgctacct ggagagacgc gcccgctgat cctttgcgaa tacgcccacg cgatgggtaa
     2101 cagtcttggc ggtttcgcta aatactggca ggcgtttcgt cagtatcccc gtttacaggg
     2161 cggcttcgtc tgggactggg tggatcagtc gctgattaaa tatgatgaaa acggcaaccc
     2221 gtggtcggct tacggcggtg attttggcga tacgccgaac gatcgccagt tctgtatgaa
     2281 cggtctggtc tttgccgacc gcacgccgca tccagcgctg acggaagcaa aacaccagca
     2341 gcagtttttc cagttccgtt tatccgggca aaccatcgaa gtgaccagcg aatacctgtt
     2401 ccgtcatagc gataacgagc tcctgcactg gatggtggcg ctggatggta agccgctggc
     2461 aagcggtgaa gtgcctctgg atgtcgctcc acaaggtaaa cagttgattg aactgcctga
     2521 actaccgcag ccggagagcg ccgggcaact ctggctcaca gtacgcgtag tgcaaccgaa
     2581 cgcgaccgca tggtcagaag ccgggcacat cagcgcctgg cagcagtggc gtctggcgga
     2641 aaacctcagt gtgacgctcc ccgccgcgtc ccacgccatc ccgcatctga ccaccagcga
     2701 aatggatttt tgcatcgagc tgggtaataa gcgttggcaa tttaaccgcc agtcaggctt
     2761 tctttcacag atgtggattg gcgataaaaa acaactgctg acgccgctgc gcgatcagtt
     2821 cacccgtgca ccgctggata acgacattgg cgtaagtgaa gcgacccgca ttgaccctaa
     2881 cgcctgggtc gaacgctgga aggcggcggg ccattaccag gccgaagcag cgttgttgca
     2941 gtgcacggca gatacacttg ctgatgcggt gctgattacg accgctcacg cgtggcagca
     3001 tcaggggaaa accttattta tcagccggaa aacctaccgg attgatggta gtggtcaaat
     3061 ggcgattacc gttgatgttg aagtggcgag cgatacaccg catccggcgc ggattggcct
     3121 gaactgccag ctggcgcagg tagcagagcg ggtaaactgg ctcggattag ggccgcaaga
     3181 aaactatccc gaccgcctta ctgccgcctg ttttgaccgc tgggatctgc cattgtcaga
     3241 catgtatacc ccgtacgtct tcccgagcga aaacggtctg cgctgcggga cgcgcgaatt
     3301 gaattatggc ccacaccagt ggcgcggcga cttccagttc aacatcagcc gctacagtca
     3361 acagcaactg atggaaacca gccatcgcca tctgctgcac gcggaagaag gcacatggct
     3421 gaatatcgac ggtttccata tggggattgg tggcgacgac tcctggagcc cgtcagtatc
     3481 ggcggaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata
     3541 ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa cgagtaatat
     3601 ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca
     3661 tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact
     3721 tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta
     3781 gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc
     3841 attaataata attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt
     3901 tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca
     3961 tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat
     4021 gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa
     4081 gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg
     4141 tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc
     4201 aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata
     4261 atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa
     4321 tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt
     4381 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4441 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4501 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     4561 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc
     4621 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4681 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4741 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4801 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4861 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4921 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4981 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5041 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5101 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5161 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5221 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5281 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5341 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5401 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5461 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5521 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5581 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5641 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5701 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5761 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5821 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5881 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5941 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     6001 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6061 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6121 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6181 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa
     6241 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6301 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6361 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6421 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6481 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6541 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6601 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6661 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6721 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6781 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagct
//
