LOCUS       sequence-49-expo        6556 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 6556)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 6556)
  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: pPD21_28
FEATURES             Location/Qualifiers
     source          1..6556
                     /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(4130..4152)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4252..4271
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4349..4367)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4435..4539
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4540..4608,4609..5400)
                     /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(4758..4777)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5571..6159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6060..6079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6313..6330
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6447..6468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6483..6488,6489..6506,6507..6513)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6521..6537
                     /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     6526..6548
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6545..6556,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6545..6556,1..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 tcatcagatc gccatctcgc gcccgtgcct
     3361 ctgacttcta agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc
     3421 accccctatt tttgttatta tcaaaaaact tctcttaatt tctttgtttt ttagcttctt
     3481 ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa tagaattaat tcgtaataaa
     3541 aagtcgaaaa aaattgtgct ccctcccccc attaataata attctatccc aaaatctaca
     3601 caatgttctg tgtacacttc ttatgttttt tacttctgat aaattttttt gaaacatcat
     3661 agaaaaaacc gcacacaaaa taccttatca tatgttacgt ttcagtttat gaccgcaatt
     3721 tttatttctt cgcacgtctg ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt
     3781 tttggagtat ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct
     3841 tttgcttttt ggggtttccc ctattgtttg tcaagatttc gaggacggcg tttttcttgc
     3901 taaaatcaca agtattgatg agcacgatgc aagaaagatc ggaagaaggt ttgggtttga
     3961 ggctcagtgg aaggtgagta gaagttgata atttgaaagt ggagtagtgt ctatggggtt
     4021 tttgccttaa atgacagaat acattcccaa tataccaaac ataactgttt cctactagtc
     4081 ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     4141 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     4201 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     4261 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4321 gaaaataccg catcaggcgg ccttaagggc ctcgtgatac gcctattttt ataggttaat
     4381 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4441 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4501 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4561 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4621 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4681 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4741 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4801 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4861 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4921 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4981 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5041 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5101 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5161 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5221 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5281 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5341 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5401 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5461 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5521 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5581 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5641 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5701 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5761 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5821 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5881 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5941 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6001 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6061 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6121 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6181 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6241 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6301 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6361 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6421 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6481 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     6541 cacacaggaa acagct
//
