LOCUS       sequence-326-exp        5343 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 5343)
  TITLE     Fire Lab C. Elegans Vector Kit 2005
REFERENCE   2  (bases 1 to 5343)
  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: pPD176.36
FEATURES             Location/Qualifiers
     source          1..5343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(148..176)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     924..944
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1046..1065)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2209..2229
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2783..2805)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2905..2924
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3002..3020)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3088..3192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3193..3261,3262..4053)
                     /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(3411..3430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4224..4812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4713..4732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4966..4983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5100..5121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5136..5141,5142..5159,5160..5166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5174..5190
                     /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     5179..5201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5198..5214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5198..5214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc ctatcgattc
       61 gcggccgctg tacacccggg tgccctatag tgagtcgtat tggtaccggt agaaaaaatg
      121 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
      181 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
      241 cttaccctta aatttatttg cactactgga aaactacctg ttccatgggt aagtttaaac
      301 atatatatac taactaaccc tgattattta aattttcagc caacacttgt cactactttc
      361 tgttatggtg ttcaatgctt ctcgagatac ccagatcata tgaaacagca tgactttttc
      421 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
      481 aactacaaga cacgtaagtt taaacagttc ggtactaact aaccatacat atttaaattt
      541 tcaggtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
      601 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca
      661 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttgt aagtttaaac
      721 atgattttac taactaacta atctgattta aattttcaga acttcaaaat tagacacaac
      781 attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat
      841 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
      901 cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca
      961 catggcatgg acgaactata caaaagccca caagctttgg aattcctgca gcccggggga
     1021 tccatggtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     1081 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     1141 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     1201 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     1261 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     1321 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     1381 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     1441 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     1501 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     1561 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     1621 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     1681 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     1741 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     1801 gagtccgggc tgcgcgttgg tgcggatgta agtttaaaca ccttaatact aactaactgt
     1861 tattatttaa attttcagat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt
     1921 tatatcccgc cgtcaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
     1981 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc
     2041 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg
     2101 ttggccgatt cattaatgca gctggcacga caggtttccc gaagcagcct gagggtaagt
     2161 ttaaacgatg aattactaac taacttgcca aatttaaatt ttcagcctcc taagaagaag
     2221 aggaaggttt gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc
     2281 tttacacttt atgcttccga cctgcagccc ggatgatcga cgccaacgtc gttgaatttt
     2341 caaattttaa atactgaata tttgtttttt ttcctattat ttatttattc tctttgtgtt
     2401 ttttttcttg ctttctaaaa aattaattca atccaaatct aaacattttt ttttctcttt
     2461 ccgtctccca attcgtattc cgctcctctc atctgaacac aatgtgcaag tttatttatc
     2521 ttctcgcttt catttcatta ggacgtgggg ggaattggtg gaagggggaa acacacaaaa
     2581 ggatgatgga aatgaaataa ggacacacaa tatgcaacaa cattcaattc agaaatatgg
     2641 aggaaggttt aaaagaaaac ataaaaatat atagaggagg aaggaaaact agtaaaaaat
     2701 aagcaaagaa attaggcgaa cgatgagaat tgtcctcgct tgggcccttt cgtctcgcgc
     2761 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2821 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2881 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2941 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cggccttaag
     3001 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3061 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3121 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3181 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3241 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3301 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3361 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3421 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3481 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3541 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3601 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3661 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3721 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3781 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3841 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3901 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3961 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4021 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4081 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4141 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4201 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4261 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4321 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4381 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4441 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4501 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4561 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     4621 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4681 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4741 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4801 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4861 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4921 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4981 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     5041 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     5101 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     5161 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     5221 tacgccaagc tgtaagttta aacatgatct tactaactaa ctattctcat ttaaattttc
     5281 agagcttaaa aatggctgaa atcactcaca acgatggata cgctaacaac ttggaaatga
     5341 aat
//
