LOCUS       sequence-323-exp        5479 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 5479)
  TITLE     Fire Lab C. Elegans Vector Kit 2005
REFERENCE   2  (bases 1 to 5479)
  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: pPD177.66
FEATURES             Location/Qualifiers
     source          1..5479
                     /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(1467..1486)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2630..2650
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        3224..3328
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3329..3397,3398..4189)
                     /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(3547..3566)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4360..4948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4849..4868
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5102..5119
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5236..5257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5272..5277,5278..5295,5296..5302)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5310..5326
                     /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     5315..5337
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5334..5350
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5334..5350
                     /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 ggttatggtc ttcaatgctt cgccagatac 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 ctgtcctacc aatctgccct
     1261 ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc
     1321 tgggattaca catggcatgg atgaactata caaatagcat tcgtagaatt gtaagtttaa
     1381 actttctcat actaactaac gcgctctatt taaattttca gaattcctgc agcccggggg
     1441 atccatggtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     1501 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     1561 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     1621 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     1681 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     1741 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     1801 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     1861 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     1921 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     1981 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     2041 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     2101 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     2161 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     2221 cgagtccggg ctgcgcgttg gtgcggatgt aagtttaaac accttaatac taactaactg
     2281 ttattattta aattttcaga tctcggtagt gggatacgac gataccgaag acagctcatg
     2341 ttatatcccg ccgtcaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     2401 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     2461 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     2521 gttggccgat tcattaatgc agctggcacg acaggtttcc cgaagcagcc tgagggtaag
     2581 tttaaacgat gaattactaa ctaacttgcc aaatttaaat tttcagcctc ctaagaagaa
     2641 gaggaaggtt tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg
     2701 ctttacactt tatgcttccg acctgcagcc cggatgatcg acgccaacgt cgttgaattt
     2761 tcaaatttta aatactgaat atttgttttt tttcctatta tttatttatt ctctttgtgt
     2821 tttttttctt gctttctaaa aaattaattc aatccaaatc taaacatttt tttttctctt
     2881 tccgtctccc aattcgtatt ccgctcctct catctgaaca caatgtgcaa gtttatttat
     2941 cttctcgctt tcatttcatt aggacgtggg gggaattggt ggaaggggga aacacacaaa
     3001 aggatgatgg aaatgaaata aggacacaca atatgcaaca acattcaatt cagaaatatg
     3061 gaggaaggtt taaaagaaaa cataaaaata tatagaggag gaaggaaaac tagtaaaaaa
     3121 taagcaaaga aattaggcga acgatgagaa ttgtcctcgc ttgggcccaa aaggcctttt
     3181 ttttttggcg ccgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3241 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3301 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3361 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3421 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3481 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3541 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3601 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3661 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3721 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3781 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3841 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3901 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3961 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4021 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4081 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4141 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4201 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4261 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4321 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4381 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4441 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4501 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4561 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4621 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4681 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4741 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4801 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4861 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4921 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4981 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5041 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5101 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5161 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5221 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     5281 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5341 cagctatgac catgattacg ccaagctgta agtttaaaca tgatcttact aactaactat
     5401 tctcatttaa attttcagag cttaaaaatg gctgaaatca ctcacaacga tggatacgct
     5461 aacaacttgg aaatgaaat
//
