LOCUS       sequence-61-expo        7024 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 7024)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7024)
  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: pPD50_21
FEATURES             Location/Qualifiers
     source          1..7024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             583..603
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             691..3738
                     /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(693..715)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4598..4620)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4720..4739
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4817..4835)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4903..5007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5008..5076,5077..5868)
                     /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(5226..5245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6039..6627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6528..6547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6781..6798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6915..6936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6951..6956,6957..6974,6975..6981)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6989..7005
                     /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     6994..7016
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7013..7024,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7013..7024,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcaccaaaaa
       61 cggaacgttg agctggacgg aaatagtggt aaagtgacat gattatagtt tgaagatttc
      121 taatttcaca attagagcaa atgttgttcg gtatttattt tcaacggtat ttatactatt
      181 ttccaccttt ttctagaaca ttcgagctgc ttgttgcaaa aggagggcga ctcacattcg
      241 gtacatggaa aagtagtgta cacaataaag agacccagat acattttccg tctgcgtctc
      301 tttgcaccca ccgggagtat tttcaaacga atgcatctag gaccttctag aacattctgt
      361 aaggctgcag aatgcgggta tataaggaaa gcgggctcag aggaagccaa cacgctttgt
      421 tctagtgcat ctaaaaaact tcgaaaatcc tcatcggatc cccgggattg gccaaaggac
      481 ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg
      541 ctagaggatc cccgggtacc gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag
      601 gtaccggtgg gtgaagacca gaaacagcac ctcgaactga gccgcgatat tgcccagcgt
      661 ttcaacgcgc tgtatggcga gatcgatccc gtcgttttac aacgtcgtga ctgggaaaac
      721 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      781 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      841 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt
      901 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc
      961 atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat
     1021 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag
     1081 acgcgaatta tttttgatgg cgttaactcg gcgtttcatc tgtggtgcaa cgggcgctgg
     1141 gtcggttacg gccaggacag tcgtttgccg tctgaatttg acctgagcgc atttttacgc
     1201 gccggagaaa accgcctcgc ggtgatggtg ctgcgctgga gtgacggcag ttatctggaa
     1261 gatcaggata tgtggcggat gagcggcatt ttccgtgacg tctcgttgct gcataaaccg
     1321 actacacaaa tcagcgattt ccatgttgcc actcgcttta atgatgattt cagccgcgct
     1381 gtactggagg ctgaagttca gatgtgcggc gagttgcgtg actacctacg ggtaacagtt
     1441 tctttatggc agggtgaaac gcaggtcgcc agcggcaccg cgcctttcgg cggtgaaatt
     1501 atcgatgagc gtggtggtta tgccgatcgc gtcacactac gtctgaacgt cgaaaacccg
     1561 aaactgtgga gcgccgaaat cccgaatctc tatcgtgcgg tggttgaact gcacaccgcc
     1621 gacggcacgc tgattgaagc agaagcctgc gatgtcggtt tccgcgaggt gcggattgaa
     1681 aatggtctgc tgctgctgaa cggcaagccg ttgctgattc gaggcgttaa ccgtcacgag
     1741 catcatcctc tgcatggtca ggtcatggat gagcagacga tggtgcagga tatcctgctg
     1801 atgaagcaga acaactttaa cgccgtgcgc tgttcgcatt atccgaacca tccgctgtgg
     1861 tacacgctgt gcgaccgcta cggcctgtat gtggtggatg aagccaatat tgaaacccac
     1921 ggcatggtgc caatgaatcg tctgaccgat gatccgcgct ggctaccggc gatgagcgaa
     1981 cgcgtaacgc gaatggtgca gcgcgatcgt aatcacccga gtgtgatcat ctggtcgctg
     2041 gggaatgaat caggccacgg cgctaatcac gacgcgctgt atcgctggat caaatctgtc
     2101 gatccttccc gcccggtgca gtatgaaggc ggcggagccg acaccacggc caccgatatt
     2161 atttgcccga tgtacgcgcg cgtggatgaa gaccagccct tcccggctgt gccgaaatgg
     2221 tccatcaaaa aatggctttc gctacctgga gagacgcgcc cgctgatcct ttgcgaatac
     2281 gcccacgcga tgggtaacag tcttggcggt ttcgctaaat actggcaggc gtttcgtcag
     2341 tatccccgtt tacagggcgg cttcgtctgg gactgggtgg atcagtcgct gattaaatat
     2401 gatgaaaacg gcaacccgtg gtcggcttac ggcggtgatt ttggcgatac gccgaacgat
     2461 cgccagttct gtatgaacgg tctggtcttt gccgaccgca cgccgcatcc agcgctgacg
     2521 gaagcaaaac accagcagca gtttttccag ttccgtttat ccgggcaaac catcgaagtg
     2581 accagcgaat acctgttccg tcatagcgat aacgagctcc tgcactggat ggtggcgctg
     2641 gatggtaagc cgctggcaag cggtgaagtg cctctggatg tcgctccaca aggtaaacag
     2701 ttgattgaac tgcctgaact accgcagccg gagagcgccg ggcaactctg gctcacagta
     2761 cgcgtagtgc aaccgaacgc gaccgcatgg tcagaagccg ggcacatcag cgcctggcag
     2821 cagtggcgtc tggcggaaaa cctcagtgtg acgctccccg ccgcgtccca cgccatcccg
     2881 catctgacca ccagcgaaat ggatttttgc atcgagctgg gtaataagcg ttggcaattt
     2941 aaccgccagt caggctttct ttcacagatg tggattggcg ataaaaaaca actgctgacg
     3001 ccgctgcgcg atcagttcac ccgtgcaccg ctggataacg acattggcgt aagtgaagcg
     3061 acccgcattg accctaacgc ctgggtcgaa cgctggaagg cggcgggcca ttaccaggcc
     3121 gaagcagcgt tgttgcagtg cacggcagat acacttgctg atgcggtgct gattacgacc
     3181 gctcacgcgt ggcagcatca ggggaaaacc ttatttatca gccggaaaac ctaccggatt
     3241 gatggtagtg gtcaaatggc gattaccgtt gatgttgaag tggcgagcga tacaccgcat
     3301 ccggcgcgga ttggcctgaa ctgccagctg gcgcaggtag cagagcgggt aaactggctc
     3361 ggattagggc cgcaagaaaa ctatcccgac cgccttactg ccgcctgttt tgaccgctgg
     3421 gatctgccat tgtcagacat gtataccccg tacgtcttcc cgagcgaaaa cggtctgcgc
     3481 tgcgggacgc gcgaattgaa ttatggccca caccagtggc gcggcgactt ccagttcaac
     3541 atcagccgct acagtcaaca gcaactgatg gaaaccagcc atcgccatct gctgcacgcg
     3601 gaagaaggca catggctgaa tatcgacggt ttccatatgg ggattggtgg cgacgactcc
     3661 tggagcccgt cagtatcggc ggaattccaa ctgagcgccg gtcgctacca ttaccaactt
     3721 gtctggtgtc aaaaataata ggcctactag cgtcgacggt accatggtat tgatatctga
     3781 gctccgcatc ggccgctgtc atcagatcgc catctcgcgc ccgtgcctct gacttctaag
     3841 tccaattact cttcaacatc cctacatgct ctttctccct gtgctcccac cccctatttt
     3901 tgttattatc aaaaaacttc tcttaatttc tttgtttttt agcttctttt aagtcacctc
     3961 taacaatgaa attgtgtaga ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa
     4021 attgtgctcc ctccccccat taataataat tctatcccaa aatctacaca atgttctgtg
     4081 tacacttctt atgtttttta cttctgataa atttttttga aacatcatag aaaaaaccgc
     4141 acacaaaata ccttatcata tgttacgttt cagtttatga ccgcaatttt tatttcttcg
     4201 cacgtctggg cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt tggagtattt
     4261 ttggaatttt tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt tgctttttgg
     4321 ggtttcccct attgtttgtc aagatttcga ggacggcgtt tttcttgcta aaatcacaag
     4381 tattgatgag cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg ctcagtggaa
     4441 ggtgagtaga agttgataat ttgaaagtgg agtagtgtct atggggtttt tgccttaaat
     4501 gacagaatac attcccaata taccaaacat aactgtttcc tactagtcgg ccgtacgggc
     4561 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4621 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4681 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     4741 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4801 tcaggcggcc ttaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4861 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4921 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4981 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5041 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5101 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5161 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5221 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5281 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5341 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5401 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5461 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5521 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5581 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5641 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5701 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5761 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5821 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5881 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5941 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6001 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6061 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6181 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6421 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6721 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6781 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6841 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6901 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6961 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7021 agct
//
