LOCUS       sequence-298-exp        4536 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 4536)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4536)
  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: pBK125
FEATURES             Location/Qualifiers
     source          1..4536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             484..504
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             517..1233
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(547..575)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1170..1190
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2110..2132)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2232..2251
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2329..2347)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2415..2519
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2520..2588,2589..3380)
                     /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(2738..2757)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3551..4139
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4040..4059
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4293..4310
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4427..4448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4463..4468,4469..4486,4487..4493)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4501..4517
                     /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     4506..4528
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4525..4536,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4525..4536,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcaccaaaaa
       61 cggaacgttg agctggacgg aaatagtggt aaagtgacat gattatagtt tgaagatttc
      121 taatttcaca attagagcaa atgttgttcg gtatttattt tcaacggtat ttatactatt
      181 ttccaccttt ttctagaaca ttcgagctgc ttgttgcaaa aggagggcga ctcacattcg
      241 gtacatggaa aagtagtgta cacaataaag agacccagat acattttccg tctgcgtctc
      301 tttgcaccca ccgggagtat tttcaaacga atgcatctag gaccttctag aacattctgt
      361 aaggctgcag aatgcgggta tataaggaaa gcgggctcag aggaagccaa cacgctttgt
      421 tctagtgcat ctaaaaaact tcgaaaatcc tcatcgggat ccccctcaga aaaaatgact
      481 gctccaaaga agaagcgtaa ggtaccggta gaaaaaatga gtaaaggaga agaacttttc
      541 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
      601 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc
      661 actactggaa aactacctgt tccatggcca acacttgtca ctactttctg ttatggtgtt
      721 caatgcttct cgagataccc agatcatatg aaacggcatg actttttcaa gagtgccatg
      781 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
      841 cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt
      901 gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
      961 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga
     1021 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     1081 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg
     1141 aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
     1201 attacacatg gcatggatga actatacaaa tagcattcgt agaattccaa ctgagcgccg
     1261 gtcgctacca ttaccaactt gtctggtgtc aaaaataata ggggccgctg tcatcagatc
     1321 gccatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca tccctacatg
     1381 ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact tctcttaatt
     1441 tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta gattcaaaaa
     1501 tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc attaataata
     1561 attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt tacttctgat
     1621 aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca tatgttacgt
     1681 ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat gacgtcaaat
     1741 catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa gtgaaagttt
     1801 atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg tcaagatttc
     1861 gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc aagaaagatc
     1921 ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata atttgaaagt
     1981 ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa tataccaaac
     2041 ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga
     2101 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     2161 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     2221 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     2281 accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc ctcgtgatac
     2341 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2401 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2461 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2521 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2581 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2641 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2701 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2761 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2821 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2881 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2941 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3001 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3061 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3121 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3181 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3241 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3301 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3361 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3421 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3481 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3541 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3601 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3661 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3721 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3781 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3841 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3901 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     3961 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4021 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4081 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4141 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4201 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4261 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4321 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     4381 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     4441 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     4501 ttgtgagcgg ataacaattt cacacaggaa acagct
//
