LOCUS       sequence-319-exp        8394 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 8394)
  TITLE     Fire Lab C. Elegans Vector Kit 2005
REFERENCE   2  (bases 1 to 8394)
  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: pPD157.99
FEATURES             Location/Qualifiers
     source          1..8394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2505..2533)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3281..3301
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5834..5856)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5956..5975
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6053..6071)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6139..6243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6244..6312,6313..7104)
                     /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(6462..6481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7275..7863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7764..7783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8017..8034
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8151..8172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8187..8192,8193..8210,8211..8217)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8225..8241
                     /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     8230..8252
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8249..8265
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8249..8265
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcaggtcgg ctataataag ttcttgaata aaataatttt cccgacaaaa
       61 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      121 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      181 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      241 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      301 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      361 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      421 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      481 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      541 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      601 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      661 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      721 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      781 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      841 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
      901 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
      961 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1021 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1081 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1141 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1201 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1261 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1321 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1381 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1441 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1501 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1561 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1621 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1681 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1741 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1801 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1861 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     1921 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     1981 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2041 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2101 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2161 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2221 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2281 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2341 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2401 gaggatccta tcgattcgcg gccgctgtac acccgggtgc cctatagtga gtcgtattgg
     2461 taccggtaga aaaaatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg
     2521 ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg
     2581 atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc
     2641 catgggtaag tttaaacata tatatactaa ctaaccctga ttatttaaat tttcagccaa
     2701 cacttgtcac tactttctgt tatggtgttc aatgcttctc gagataccca gatcatatga
     2761 aacagcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat
     2821 ttttcaaaga tgacgggaac tacaagacac gtaagtttaa acagttcggt actaactaac
     2881 catacatatt taaattttca ggtgctgaag tcaagtttga aggtgatacc cttgttaata
     2941 gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg
     3001 aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca
     3061 aagttgtaag tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagaact
     3121 tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa
     3181 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat
     3241 ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa
     3301 cagctgctgg gattacacat ggcatggacg aactatacaa aagcccacaa gctttggaat
     3361 tcgctagcaa accggtcacc atgcaaaagt cagatgaagt cactgaaaag ttcaaacgat
     3421 attgtaatca actggaaaaa tacggccaaa ctgaaaatgt gcattctcca gtgatggcga
     3481 tgttgcgcag aaagtatgtt ttattctgta acaaaaactt aaaatacaac agcttgagtc
     3541 agaaactatg aaattaataa acgatcttta agtttaaaat ttaaagaatt acaattttca
     3601 gaggccgaaa acagctcatt gagattatga aacgcgacgg ggactgtaca agttcaatca
     3661 ataaattgtg gattgttgga tattatcacc ctttccagtt ttttattcga gtaagattaa
     3721 cttttagtca ttgtattatt tttatttgat ttccaaaggt tcttattaca ggataaggag
     3781 aaaaacatgg caatagccgt gttgctgaca atgttctgtg gagagcttca agaaatgctt
     3841 tcgcttcccg atgacaaata cccagcattg tggaacatgt acattggaga ttttcatcgt
     3901 tatatgccag acgaagaaat tcagaaatgc ttggcagttg gatattattc aagagcaatc
     3961 gatctagatc cgaatcaagg aagagctttc cacgttctcg ctggtctccg agctgacctg
     4021 aatgttgcac agaagctgag actgatgatt ttaggacagc tcgccgatgc tccatataaa
     4081 aaaggcactg aactcttgga atatttgaaa tttccacaaa aagaatccac tgataaattg
     4141 atggtagatt ttgtcatttg ggcattgaac gagaaatcaa aacgaatgga ttatcagatg
     4201 actggaatca aaatagtaaa cgagtttaaa gcagaaatcg aacagaaact cgaattcgat
     4261 tggtctctga tcatgtcaac gtgtcgttta gcatctaaat tggcaatgaa gaaatttgga
     4321 tgtaagtacc gattccaata tctatagaaa attcttattc tgctgtagga taacataatt
     4381 ttttcagttc aacaattcta caattgcttc gacacaattt caacgctgta tattacaatt
     4441 tattcacgta cgatttcatc caaatgcctt ctagctgaag ctatctcatg gatttccgat
     4501 tcagccgaaa tactcggaca cttggatgag cagaaaaatg agccacattt ccagaagcta
     4561 tcggttttcg ccaaaacggt aagatttaga gataagaaaa tgtagaatag atgtttcaat
     4621 tttaaaatca aggggaatta attataaata taacttattt tcagaaatgg aacgagctca
     4681 acgacttggt tatgaatcat atcaattccg tattcacatc catgtcactc acaatcaatc
     4741 catctatttc tatgaccagc tttctgctga atggtccaat atccgaaccc aatgttgaat
     4801 ttttgagtca attaatcaat tatctggttt ccgtcgaatt cccaccaatg gagataatcc
     4861 acgatagaga agaatcgggt ccattgctac gacgaattaa tcaatcggaa caaaaacgtt
     4921 tggatattca aataaagaca caaaatgatg aagttaaccg ggaggattgg agacctgttt
     4981 atgtattgat ggattatgac gtcatagttg acaaaataag aattgctcta aaaatttggg
     5041 atatcgatga tttcatctgc attttgccat caacagtgct tgatgaactt gattatcaga
     5101 aaacgaagaa tcgagcagtt cgtccagtaa ttcgggcatt aatggagctt caagcagagg
     5161 gaaaaattgt gctaaagaaa tgtgataacg agagaagctg cgcggagcaa ttggttcaaa
     5221 gtgcacgaag atctgcagaa gatcacaagc atattgtcgc ttttctgtgc aaaaacccga
     5281 gtgaacagaa agaaatggaa ggagttactt tttacgatat cgatcagttc tacatgaagt
     5341 atttggagtg acctaggttc cggatgatcg acgccaacgt cgttgaattt tcaaatttta
     5401 aatactgaat atttgttttt tttcctatta tttatttatt ctctttgtgt tttttttctt
     5461 gctttctaaa aaattaattc aatccaaatc taaacatttt tttttctctt tccgtctccc
     5521 aattcgtatt ccgctcctct catctgaaca caatgtgcaa gtttatttat cttctcgctt
     5581 tcatttcatt aggacgtggg gggaattggt ggaaggggga aacacacaaa aggatgatgg
     5641 aaatgaaata aggacacaca atatgcaaca acattcaatt cagaaatatg gaggaaggtt
     5701 taaaagaaaa cataaaaata tatagaggag gaaggaaaac tagtaaaaaa taagcaaaga
     5761 aattaggcga acgatgagaa ttgtcctcgc ttgggccctt tcgtctcgcg cgtttcggtg
     5821 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     5881 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5941 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6001 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     6061 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     6121 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6181 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6241 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6301 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6361 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6421 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6481 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6541 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6601 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6661 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6721 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6781 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6841 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6901 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6961 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7021 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7081 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7141 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7201 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7261 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7321 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7381 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     7441 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7501 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7561 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7621 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     7681 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7741 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7801 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7861 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7921 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7981 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     8041 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     8101 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     8161 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     8221 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     8281 ctgtaagttt aaacatgatc ttactaacta actattctca tttaaatttt cagagcttaa
     8341 aaatggctga aatcactcac aacgatggat acgctaacaa cttggaaatg aaat
//
