LOCUS       sequence-322-exp        5428 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 5428)
  TITLE     Fire Lab C. Elegans Vector Kit 2005
REFERENCE   2  (bases 1 to 5428)
  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: pPD176.86
FEATURES             Location/Qualifiers
     source          1..5428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(518..546)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1294..1314
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1416..1435)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2579..2599
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        3173..3277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3278..3346,3347..4138)
                     /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(3496..3515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4309..4897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4798..4817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5051..5068
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5185..5206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5221..5226,5227..5244,5245..5251)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5259..5275
                     /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     5264..5286
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5283..5299
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5283..5299
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc cccgggattg
       61 gccaaaggac ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga
      121 ggacccttgg agggtaccta atttgatcga taatatgtac atattttgtt attcgttccg
      181 gtatcgtacc tccacctacc ataaatccca taccctacat ctttatcgat tttttgcaaa
      241 ttacgagcgt tgtagggggc ggagcgatag gtcctatagg ttttggtata tcatcattca
      301 ttcattcatt ggtacattca tttacccacc ttcctctttc tgagcttctc tggagttctg
      361 tgcttccttt ttcccttatc tttatactgt aatttttaac tttcaggcat tgattggatc
      421 ctatcgattc gcggccgctg tacacccggg tgccctatag tgagtcgtat tggtaccggt
      481 agaaaaaatg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
      541 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
      601 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatgggt
      661 aagtttaaac atatatatac taactaaccc tgattattta aattttcagc caacacttgt
      721 cactactttc tgttatggtg ttcaatgctt ctcgagatac ccagatcata tgaaacagca
      781 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa
      841 agatgacggg aactacaaga cacgtaagtt taaacagttc ggtactaact aaccatacat
      901 atttaaattt tcaggtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga
      961 gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa
     1021 ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttgt
     1081 aagtttaaac atgattttac taactaacta atctgattta aattttcaga acttcaaaat
     1141 tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc
     1201 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct
     1261 ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc
     1321 tgggattaca catggcatgg acgaactata caaaagccca caagctttgg aattcctgca
     1381 gcccggggga tccatggtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     1441 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     1501 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     1561 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     1621 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     1681 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     1741 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     1801 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     1861 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     1921 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     1981 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     2041 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     2101 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     2161 cgccattacc gagtccgggc tgcgcgttgg tgcggatgta agtttaaaca ccttaatact
     2221 aactaactgt tattatttaa attttcagat ctcggtagtg ggatacgacg ataccgaaga
     2281 cagctcatgt tatatcccgc cgtcaaccac catcaaacag gattttcgcc tgctggggca
     2341 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     2401 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     2461 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gaagcagcct
     2521 gagggtaagt ttaaacgatg aattactaac taacttgcca aatttaaatt ttcagcctcc
     2581 taagaagaag aggaaggttt gagcgcaacg caattaatgt aagttagctc actcattagg
     2641 caccccaggc tttacacttt atgcttccga cctgcagccc ggatgatcga cgccaacgtc
     2701 gttgaatttt caaattttaa atactgaata tttgtttttt ttcctattat ttatttattc
     2761 tctttgtgtt ttttttcttg ctttctaaaa aattaattca atccaaatct aaacattttt
     2821 ttttctcttt ccgtctccca attcgtattc cgctcctctc atctgaacac aatgtgcaag
     2881 tttatttatc ttctcgcttt catttcatta ggacgtgggg ggaattggtg gaagggggaa
     2941 acacacaaaa ggatgatgga aatgaaataa ggacacacaa tatgcaacaa cattcaattc
     3001 agaaatatgg aggaaggttt aaaagaaaac ataaaaatat atagaggagg aaggaaaact
     3061 agtaaaaaat aagcaaagaa attaggcgaa cgatgagaat tgtcctcgct tgggcccaaa
     3121 aggccttttt tttttggcgc cgacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3181 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3241 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3301 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3361 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3421 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3481 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3541 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3601 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3661 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3721 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3781 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3841 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3901 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3961 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4021 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4081 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4141 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4201 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4261 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4321 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4381 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4441 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4501 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4561 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4621 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4681 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4741 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4801 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4861 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4921 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4981 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5041 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     5101 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5161 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5221 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5281 cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat gatcttacta
     5341 actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac tcacaacgat
     5401 ggatacgcta acaacttgga aatgaaat
//
