LOCUS       sequence-307-exp        5108 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 5108)
  TITLE     Fire Lab C. Elegans Vector Kit 2005
REFERENCE   2  (bases 1 to 5108)
  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: pPD177.62
FEATURES             Location/Qualifiers
     source          1..5108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             503..523
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(597..616)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(1277..1305)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2053..2073
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2682..2704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2804..2823
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2901..2919)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2987..3091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3092..3160,3161..3952)
                     /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(3310..3329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4123..4711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4612..4631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4865..4882
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4999..5020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5035..5040,5041..5058,5059..5065)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5073..5089
                     /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     5078..5100
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5097..5108,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5097..5108,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta
       61 cctaatttga tcgataatat gtacatattt tgttattcgt tccggtatcg tacctccacc
      121 taccataaat cccataccct acatctttat cgattttttg caaattacga gcgttgtagg
      181 gggcggagcg ataggtccta taggttttgg tatatcatca ttcattcatt cattggtaca
      241 ttcatttacc caccttcctc tttctgagct tctctggagt tctgtgcttc ctttttccct
      301 tatctttata ctgtaatttt taactttcag gcattgattg gatccccggg attggccaaa
      361 ggacccaaag gtatgtttcg aatgatacta acataacata gaacattttc aggaggaccc
      421 ttggctagag gatccccggg attggccaaa ggacccaaag gaggaccctt ggagggtacc
      481 gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag gtaccggcta gattagataa
      541 aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac
      601 aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa
      661 aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc accatactca
      721 cttttgccct ttagaagggg aaagctggca agatttttta cgtaagttta aacatattgt
      781 tactaactaa ccttatacat ttaaattttc aggtaataac gctaaaagtt ttagatgtgc
      841 tttactaagt catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca
      901 gtatgaaact ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa
      961 tgcattatat gcactcagcg ctgtggggca ttttacttta ggttgcgtat tggaagatca
     1021 agagcatcaa gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt
     1081 attacgacaa gctatcgaat tatttgatca ccagtaagtt taaacgtgga gatactaact
     1141 aaccggcagt atttaaattt tcagaggtgc agagccagcc ttcttattcg gccttgaatt
     1201 gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtctacta gaaaaggaga
     1261 agaactcttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
     1321 caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa
     1381 atttatttgc actactggaa aactacctgt tccatgggta agtttaaaca tatatatact
     1441 aactaaccct gattatttaa attttcagcc aacacttgtc actactttct catggggcgt
     1501 tcaatgcttc tcgagatacc cagatcatat gaaacagcat gactttttca agagtgccat
     1561 gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac
     1621 acgtaagttt aaacagttcg gtactaacta accatacata tttaaatttt caggtgctga
     1681 agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa
     1741 agaagatgga aacattcttg gacacaaatt ggaatacaac tatatttcac acaacgtata
     1801 catcactgcc gacaaacaaa agaatgggat caaagcggta agtttaaaca tgattttact
     1861 aactaactaa tctgatttaa attttcagaa cttcaaaatt agacacaaca ttgaagatgg
     1921 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     1981 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     2041 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     2101 tgaactatac aaatagcatt cgtagaattg taagtttaaa ctttctcata ctaactaacg
     2161 cgctctattt aaattttcag aattcgctag ccggccatac aagtaatccg gatgatcgac
     2221 gccaacgtcg ttgaattttc aaattttaaa tactgaatat ttgttttttt tcctattatt
     2281 tatttattct ctttgtgttt tttttcttgc tttctaaaaa attaattcaa tccaaatcta
     2341 aacatttttt tttctctttc cgtctcccaa ttcgtattcc gctcctctca tctgaacaca
     2401 atgtgcaagt ttatttatct tctcgctttc atttcattag gacgtggggg gaattggtgg
     2461 aagggggaaa cacacaaaag gatgatggaa atgaaataag gacacacaat atgcaacaac
     2521 attcaattca gaaatatgga ggaaggttta aaagaaaaca taaaaatata tagaggagga
     2581 aggaaaacta gtaaaaaata agcaaagaaa ttaggcgaac gatgagaatt gtcctcgctt
     2641 gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2701 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2761 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     2821 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2881 cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2941 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3001 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3061 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3121 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3181 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3241 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3301 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3361 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3421 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3481 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3541 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3601 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3661 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3721 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3781 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3841 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3901 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3961 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4021 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4081 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4141 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4201 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4261 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4321 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4381 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4441 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4501 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4561 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4621 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4681 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4741 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4801 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4861 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     4921 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     4981 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     5041 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     5101 aaacagct
//
