LOCUS       sequence-67-expo        8984 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 8984)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 8984)
  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: pPD80_08
FEATURES             Location/Qualifiers
     source          1..8984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1343..1363
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1376..2089
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1406..1434)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2029..2049
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2180..5227
                     /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(2182..2204)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6558..6580)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6680..6699
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6777..6795)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6863..6967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6968..7036,7037..7828)
                     /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(7186..7205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7999..8587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8488..8507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8741..8758
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8875..8896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8911..8916,8917..8934,8935..8941)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8949..8965
                     /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     8954..8976
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8973..8984,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8973..8984,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccagctt
       61 gcatgcctgc aggtcgaggc atttgaattg ggggtggtgg acagtaactg tctgtaataa
      121 taattactcc tgaccaggtt gcaattcgag ttttgataag cataattata ccttgtacat
      181 tgtgggtttt gtgctgtgga cgttttattg tgacatcccc ataagctaca agaacaaaaa
      241 tgaaatgtaa aagtattgaa aacgtcgtac attttatatc tgagtagtat cctttgcttt
      301 aaatgtccat aaaactaatt ttataatcaa taaaacaacg tttgtaaatc aactgagttt
      361 acaagtagag acattgaggg atactttcac tatgctaaag tgataatcga ccaaataata
      421 acttcacttt ggtatttatt cctgtcttat aaatgttatg tatgaattaa attcatatgc
      481 atatggctca ctctgacaat ttttaataat cttccagatc aatattgact accgatgcgg
      541 gtggtctttt gctttgaatt ctgctgaact ttacaccccg aacagcaatg tgtgcttcag
      601 cctaaaaaaa agtaagtgtg ttaatcagtg cccccgattc ttcatttttt gcccctctct
      661 cccgtttcgt cggcaaaaga agagaaaata aagataagtc tcaagatagg ttggtaatcg
      721 ctaaagtggt tgtgtggata agagtagcaa aatggcagga agagcacttt gcgcgcacac
      781 actgtactca ttgttctgga taaaattctc tcgttgtttg ccgtcggatg tctgcctctc
      841 tgcattgagc cggcttcttc actatcttta gttaacctaa aatgccgttt cttttctcgt
      901 atccccacta tcccgttgag gttctctgct ctcttcgctc cctaccgcca gcgagcaact
      961 atccgtgggg gcgccttgct cggaagatgg gggggaagaa agaagatttt tgctatttgc
     1021 acttgagaaa gagacttttc ctgcgtcgat ggttagagaa cagtgtgcag acacttttca
     1081 gctacctaga attacaattg gatatccccg cctcccaatc cacccaccca gggaaaaaga
     1141 agggctcgcc gaaaatcaaa gttatctcca ggctcgcgca tcccaccgag cggttgactt
     1201 ctctccacca cttttcattt taaccctcgg ggtacgggat tggccaaagg acccaaaggt
     1261 atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt ggagggtacc
     1321 gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag gtaccggtag aaaaaatgag
     1381 taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
     1441 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact
     1501 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac
     1561 tactttctct tatggtgttc aatgcttttc aagataccca gatcatatga aacggcatga
     1621 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga
     1681 tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag
     1741 aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga
     1801 atacaactat aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa
     1861 agttaacttc aaaattagac acaacattga agatggaagc gttcaactag cagaccatta
     1921 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
     1981 cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga
     2041 gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaat gcccggtggg
     2101 tgaagaccag aaacagcacc tcgaactgag ccgcgatatt gcccagcgtt tcaacgcgct
     2161 gtatggcgag atcgatcccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2221 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2281 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg
     2341 gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga
     2401 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa
     2461 cgtgacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg
     2521 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat
     2581 ttttgatggc gttaactcgg cgtttcatct gtggtgcaac gggcgctggg tcggttacgg
     2641 ccaggacagt cgtttgccgt ctgaatttga cctgagcgca tttttacgcg ccggagaaaa
     2701 ccgcctcgcg gtgatggtgc tgcgctggag tgacggcagt tatctggaag atcaggatat
     2761 gtggcggatg agcggcattt tccgtgacgt ctcgttgctg cataaaccga ctacacaaat
     2821 cagcgatttc catgttgcca ctcgctttaa tgatgatttc agccgcgctg tactggaggc
     2881 tgaagttcag atgtgcggcg agttgcgtga ctacctacgg gtaacagttt ctttatggca
     2941 gggtgaaacg caggtcgcca gcggcaccgc gcctttcggc ggtgaaatta tcgatgagcg
     3001 tggtggttat gccgatcgcg tcacactacg tctgaacgtc gaaaacccga aactgtggag
     3061 cgccgaaatc ccgaatctct atcgtgcggt ggttgaactg cacaccgccg acggcacgct
     3121 gattgaagca gaagcctgcg atgtcggttt ccgcgaggtg cggattgaaa atggtctgct
     3181 gctgctgaac ggcaagccgt tgctgattcg aggcgttaac cgtcacgagc atcatcctct
     3241 gcatggtcag gtcatggatg agcagacgat ggtgcaggat atcctgctga tgaagcagaa
     3301 caactttaac gccgtgcgct gttcgcatta tccgaaccat ccgctgtggt acacgctgtg
     3361 cgaccgctac ggcctgtatg tggtggatga agccaatatt gaaacccacg gcatggtgcc
     3421 aatgaatcgt ctgaccgatg atccgcgctg gctaccggcg atgagcgaac gcgtaacgcg
     3481 aatggtgcag cgcgatcgta atcacccgag tgtgatcatc tggtcgctgg ggaatgaatc
     3541 aggccacggc gctaatcacg acgcgctgta tcgctggatc aaatctgtcg atccttcccg
     3601 cccggtgcag tatgaaggcg gcggagccga caccacggcc accgatatta tttgcccgat
     3661 gtacgcgcgc gtggatgaag accagccctt cccggctgtg ccgaaatggt ccatcaaaaa
     3721 atggctttcg ctacctggag agacgcgccc gctgatcctt tgcgaatacg cccacgcgat
     3781 gggtaacagt cttggcggtt tcgctaaata ctggcaggcg tttcgtcagt atccccgttt
     3841 acagggcggc ttcgtctggg actgggtgga tcagtcgctg attaaatatg atgaaaacgg
     3901 caacccgtgg tcggcttacg gcggtgattt tggcgatacg ccgaacgatc gccagttctg
     3961 tatgaacggt ctggtctttg ccgaccgcac gccgcatcca gcgctgacgg aagcaaaaca
     4021 ccagcagcag tttttccagt tccgtttatc cgggcaaacc atcgaagtga ccagcgaata
     4081 cctgttccgt catagcgata acgagctcct gcactggatg gtggcgctgg atggtaagcc
     4141 gctggcaagc ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt tgattgaact
     4201 gcctgaacta ccgcagccgg agagcgccgg gcaactctgg ctcacagtac gcgtagtgca
     4261 accgaacgcg accgcatggt cagaagccgg gcacatcagc gcctggcagc agtggcgtct
     4321 ggcggaaaac ctcagtgtga cgctccccgc cgcgtcccac gccatcccgc atctgaccac
     4381 cagcgaaatg gatttttgca tcgagctggg taataagcgt tggcaattta accgccagtc
     4441 aggctttctt tcacagatgt ggattggcga taaaaaacaa ctgctgacgc cgctgcgcga
     4501 tcagttcacc cgtgcaccgc tggataacga cattggcgta agtgaagcga cccgcattga
     4561 ccctaacgcc tgggtcgaac gctggaaggc ggcgggccat taccaggccg aagcagcgtt
     4621 gttgcagtgc acggcagata cacttgctga tgcggtgctg attacgaccg ctcacgcgtg
     4681 gcagcatcag gggaaaacct tatttatcag ccggaaaacc taccggattg atggtagtgg
     4741 tcaaatggcg attaccgttg atgttgaagt ggcgagcgat acaccgcatc cggcgcggat
     4801 tggcctgaac tgccagctgg cgcaggtagc agagcgggta aactggctcg gattagggcc
     4861 gcaagaaaac tatcccgacc gccttactgc cgcctgtttt gaccgctggg atctgccatt
     4921 gtcagacatg tataccccgt acgtcttccc gagcgaaaac ggtctgcgct gcgggacgcg
     4981 cgaattgaat tatggcccac accagtggcg cggcgacttc cagttcaaca tcagccgcta
     5041 cagtcaacag caactgatgg aaaccagcca tcgccatctg ctgcacgcgg aagaaggcac
     5101 atggctgaat atcgacggtt tccatatggg gattggtggc gacgactcct ggagcccgtc
     5161 agtatcggcg gaattccaac tgagcgccgg tcgctaccat taccaacttg tctggtgtca
     5221 aaaataatag gggccgctgt catcagatcg ccatctcgcg cccgtgcctc tgacttctaa
     5281 gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt
     5341 ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt taagtcacct
     5401 ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa
     5461 aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt
     5521 gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata gaaaaaaccg
     5581 cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc
     5641 gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt
     5701 tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg
     5761 gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct aaaatcacaa
     5821 gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag gctcagtgga
     5881 aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa
     5941 tgacagaata cattcccaat ataccaaaca taactgttta aaattaaaca tttttctaaa
     6001 ttttatatga tttcttttaa atttgcaaaa attacttaaa tttgaattcc cgcgcaaatg
     6061 agtgacttca ttttctgcat tattgtgttt tccggctata ttaataggta tttgtttgtg
     6121 tttttcttta ttttatgatt cgaactccaa tttgtaaatt ttcgaacata tttccctaaa
     6181 gaaaaaatat gattaatctg gaaaaattgg aaaattattt ttcaaataaa aaacaaagaa
     6241 aaaaatgaag aaaaacctat tagtttggcc ataaaacgca aaaatgtcga aaatgacgtc
     6301 actcatctgc gcgggaaatc aagaataatt cggccttttt tatttttttg gaaaatcgta
     6361 aaacatttag aaaaattttt taatagttat atgggactta ttctgcattt agggcaaaag
     6421 ccagagacta ctccaccgtt aacatgaatt atgaatatta ttgcgacaag acccaaacat
     6481 tgataaaccc aaatctagcc tactagtcgg ccgtacgggc cctttcgtct cgcgcgtttc
     6541 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     6601 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6661 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     6721 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct
     6781 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     6841 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6901 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6961 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     7021 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     7081 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     7141 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     7201 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7261 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7321 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7381 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7441 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7501 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7561 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7621 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7681 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7741 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7801 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7861 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7921 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7981 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     8041 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     8101 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     8161 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     8221 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     8281 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8341 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     8401 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8461 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8521 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8581 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8641 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     8701 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     8761 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     8821 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     8881 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     8941 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agct
//
