LOCUS       sequence-83-expo        4438 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 4438)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 4438)
  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: pPD91_14
FEATURES             Location/Qualifiers
     source          1..4438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             386..406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             419..1135
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(449..477)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1072..1092
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2012..2034)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2134..2153
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2231..2249)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2317..2421
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2422..2490,2491..3282)
                     /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(2640..2659)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3453..4041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3942..3961
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4195..4212
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4329..4350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4365..4370,4371..4388,4389..4395)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4403..4419
                     /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     4408..4430
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4427..4438,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4427..4438,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccaattc
       61 ttgagacttc cactatcttc tctggcaccc tccactacgc gtaggcgtct ctcgcttcgt
      121 gtattcccgg gaagccgggt tcccgtctct cccgccgctg ccgctgccgc acacagcttt
      181 acacctcgta gaatccccaa agaggggcgt ggcttgcggg tgccaacatc ctcctgccga
      241 ggaagaagca ggcactcatc actcgcatca tcaacctcgg gattggccaa aggacccaaa
      301 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      361 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tagaaaaaat
      421 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
      481 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
      541 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt
      601 cactactttc tcttatggtg ttcaatgctt ttcaagatac ccagatcata tgaaacggca
      661 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa
      721 agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa
      781 tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt
      841 ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat
      901 caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca
      961 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     1021 gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct
     1081 tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca aatagcattc
     1141 gtagaattcc aactgagcgc cggtcgctac cattaccaac ttgtctggtg tcaaaaataa
     1201 taggggccgc tgtcatcaga tcgccatctc gcgcccgtgc ctctgacttc taagtccaat
     1261 tactcttcaa catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat
     1321 tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc ttttaagtca cctctaacaa
     1381 tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg
     1441 ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact
     1501 tcttatgttt tttacttctg ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa
     1561 aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc
     1621 tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt atttttggaa
     1681 tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt ttggggtttc
     1741 ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca caagtattga
     1801 tgagcacgat gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag
     1861 tagaagttga taatttgaaa gtggagtagt gtctatgggg tttttgcctt aaatgacaga
     1921 atacattccc aatataccaa acataactgt ttcctactag tcggccgtac gggccctttc
     1981 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     2041 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     2101 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     2161 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     2221 ggccttaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     2281 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2341 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2401 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2461 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2521 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2581 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2641 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2701 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2761 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2821 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2881 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2941 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3001 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3061 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3121 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3181 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3241 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3301 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3361 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3421 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3481 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3541 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3601 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3661 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3721 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3781 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3841 gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3901 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3961 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4021 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4081 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4141 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4201 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4261 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4321 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4381 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagct
//
