LOCUS       sequence-327-exp        6581 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 6581)
  TITLE     Fire Lab C. Elegans Vector Kit 2005
REFERENCE   2  (bases 1 to 6581)
  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: pPD138.11
FEATURES             Location/Qualifiers
     source          1..6581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      16..25
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(22..24,25..27,28..738)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(22..24,25..27,28..738)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(67..88)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(328..347)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     675..696
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1683..1705)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1805..1824
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1902..1920)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1988..2092
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2093..2161,2162..2953)
                     /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(2311..2330)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3124..3712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3613..3632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3866..3883
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4000..4021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4036..4041,4042..4059,4060..4066)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4074..4090
                     /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     4079..4101
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4098..4114
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4098..4114
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
       61 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      121 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      181 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
      241 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      301 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      361 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      421 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
      481 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
      541 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      601 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
      661 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
      721 atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttcgaa ttccaactga
      781 gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat
      841 cagagtaagt ttaaactgag ttctactaac taacgagtaa tatttaaatt ttcagcatct
      901 cgcgcccgtg cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc
      961 tccctgtgct cccaccccct atttttgtta ttatcaaaaa acttctctta atttctttgt
     1021 tttttagctt cttttaagtc acctctaaca atgaaattgt gtagattcaa aaatagaatt
     1081 aattcgtaat aaaaagtcga aaaaaattgt gctccctccc cccattaata ataattctat
     1141 cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt ttttacttct gataaatttt
     1201 tttgaaacat catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt
     1261 tatgaccgca atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc
     1321 atcgtgaaaa agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat
     1381 taattttcct gcttttgctt tttggggttt cccctattgt ttgtcaagat ttcgaggacg
     1441 gcgtttttct tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa
     1501 ggtttgggtt tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag
     1561 tgtctatggg gtttttgcct taaatgacag aatacattcc caatatacca aacataactg
     1621 tttcctacta gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     1681 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     1741 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     1801 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     1861 agatgcgtaa ggagaaaata ccgcatcagg cggccttaag ggcctcgtga tacgcctatt
     1921 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     1981 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2041 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2101 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2161 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2221 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2281 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2341 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2401 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2461 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2521 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2581 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2641 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2701 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2761 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2821 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2881 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2941 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3001 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3061 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3121 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3181 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3241 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3301 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3361 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3421 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3481 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga
     3541 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3601 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3661 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3721 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3781 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3841 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     3901 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3961 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     4021 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     4081 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgggctgca
     4141 ggtcggctat aataagttct tgaataaaat aattttcccg acaaaacatg agtatttctt
     4201 tcgaaaataa aagtgcaggc taattagaga ttattctgta attaactgca taatttgtca
     4261 cgtgccatag ttttacattc cactacgtca tagttcttaa aatactaatc tcctgaaaat
     4321 agaagtaggt gaagaaagtt taattatcag ttctaaaatg acaattgatc tttggaatat
     4381 gttctgaaac taccgatcat tgaacagatg ctatttgaat gatatagaat tgtatatttg
     4441 caatttctga aacgcgttct taaaggcaca cagattaatt caaaagggtc tggccgcaaa
     4501 aaggtttatg gtggccgatt ttgagttttg tgtgtgattg ctttttcaca atcagtgttt
     4561 tcaggattat gtgatgaact agatcttcaa gtttcgttac atttcatatg ttttcggaac
     4621 tcacgaagta catattgggt attgtgctca aaaaattcag caatcagctt cgctccgctg
     4681 actttagaac ccaaaaaaat agtatggcca aactgactgt gttacgatca tttcaatttt
     4741 tcaatacata tttaagattt ctaagagtaa gaaggtcaaa aactgttctg gaatacatat
     4801 atatttttca ggttacaatt agtcaaaaag tgcactgaaa tatacgtttt aatttcacga
     4861 ataacccaat tagttcaatg tatttttggt caaccaacgt taaagtttgg cttccaacca
     4921 attatcattt ctgatcaacc acaatgtttt ttctttatct gcaagttaat tttttatttt
     4981 tatccagatg tttggcatat ttttcaattc ttcactagcg cccacttctt gcacttccgg
     5041 cgccctgaat ctaatgcatc tgttgcaaga attgaaagac caatcaacac attgttttct
     5101 tcacgagata ctgaagaaaa tgaataaaaa cagagaaaaa gagccatgtg attagtgaca
     5161 actgttgcta acagatacca tagcttggac ttggtacgtg atggcaacgt atgggtcaac
     5221 aaaaatgatt gcagaggggg tgcaaaacag tcaagtcgag aaaatatgaa aaacagaaaa
     5281 caaagaacag aaaaatgggt ttgagagtca gtataattta taaaagaaaa attgtacata
     5341 gaaattaacc atttttgtag aagaagttat ttttcaagca tcgttaaaaa ttattcaaag
     5401 caccttattt catatttaat tttaaacatg gttaaatgaa caacacggtg cgcaatcagg
     5461 aaaacttgaa atctgaaact gttgttgtga tcttcttcgc aactgttcag atagcactag
     5521 tgtaatgtta agagtgcgcg aatataatgg aatataatgg atcacacctc ctgccatcag
     5581 gtaaacgtct ctgttatcac atatttccaa ctattaaatt tttacctttt acagttttac
     5641 atttttttga aaaaagtaac tttttgtctt caaaatccct gacgaaaata tcaaatattt
     5701 taatcgagac tgcagaggaa ccgattgatg atttggaaaa tccagcttta cctgtgtaag
     5761 aactgaaaag tttcataacc ctagggtatt cccagttaca ttccccactg gctaacaata
     5821 gcacccagtt tttcatcacc tttcttcaaa tttctcggcg atttgttaaa aacaaaattt
     5881 gtgtcccttc tctgatatct ctatgtctct aaacacaagt tcatcggaaa acgaaggagg
     5941 gtaggtgttg gttgggctcc cgaagtgaaa atagaagagc aagaatagaa tattagagag
     6001 agagtgcaga gagggcggga tagctcccgg gattccgttt tcttcttctt tatcttcaac
     6061 gatgatgtgt gtgcgtgttg tatagattct gttgctcccc cacaactcgc tccgaaggct
     6121 caatacaatt caattgatat tggaggagag cctaccggag tgggaggata agaagaaaca
     6181 taagaagaag aagaagaaga agcatgcttc tggtttttga tgctatgaaa acggcacaaa
     6241 aagatgattg aggtcccttt tcaatacctt ctctcatctt tcaaatccca ttgaaaccta
     6301 aaacttctca ccacgcttta ccattgttct ccaaaaactt atagcaatgt ctataacttt
     6361 tttatctctg aaaagcagtg ttccattttt ctttttccta ttttatttca attgtttctc
     6421 acatttcgtt tggattcttt gcttgtcaac cagcttcttc ttccactttt accgtctaat
     6481 tttcagggca gggagccatc aaacccacga ccactagatc catctagagg atccccggga
     6541 ttggccaaag gacccaaagg aggacccttg gagggtaccg g
//
