LOCUS       sequence-124-exp        5740 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 5740)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 5740)
  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: L2630
FEATURES             Location/Qualifiers
     source          1..5740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1433..1453
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1496..1524)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2272..2292
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3314..3336)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3436..3455
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3533..3551)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3619..3723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3724..3792,3793..4584)
                     /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(3942..3961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4755..5343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5244..5263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5497..5514
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5631..5652
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5667..5672,5673..5690,5691..5697)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5705..5721
                     /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     5710..5732
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5729..5740,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5729..5740,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcgtcatcca
       61 cagtgtaatg taagatggtt cagagattca gaagagaggc agagatagcc tcatcccact
      121 caattgatga gagaatggga tccccagctt gcatgcctgc aggtcgaggc atttgaattg
      181 ggggtggtgg acagtaactg tctgtaataa taattactcc tgaccaggtt gcaattcgag
      241 ttttgataag cataattata ccttgtacat tgtgggtttt gtgctgtgga cgttttattg
      301 tgacatcccc ataagctaca agaacaaaaa tgaaatgtaa aagtattgaa aacgtcgtac
      361 attttatatc tgagtagtat cctttgcttt aaatgtccat aaaactaatt ttataatcaa
      421 taaaacaacg tttgtaaatc aactgagttt acaagtagag acattgaggg atactttcac
      481 tatgctaaag tgataatcga ccaaataata acttcacttt ggtatttatt cctgtcttat
      541 aaatgttatg tatgaattaa attcatatgc atatggctca ctctgacaat ttttaataat
      601 cttccagatc aatattgact accgatgcgg gtggtctttt gctttgaatt ctgctgaact
      661 ttacaccccg aacagcaatg tgtgcttcag cctaaaaaaa agtaagtgtg ttaatcagtg
      721 cccccgattc ttcatttttt gcccctctct cccgtttcgt cggcaaaaga agagaaaata
      781 aagataagtc tcaagatagg ttggtaatcg ctaaagtggt tgtgtggata agagtagcaa
      841 aatggcagga agagcacttt gcgcgcacac actgtactca ttgttctgga taaaattctc
      901 tcgttgtttg ccgtcggatg tctgcctctc tgcattgagc cggcttcttc actatcttta
      961 gttaacctaa aatgccgttt cttttctcgt atccccacta tcccgttgag gttctctgct
     1021 ctcttcgctc cctaccgcca gcgagcaact atccgtgggg gcgccttgct cggaagatgg
     1081 gggggaagaa agaagatttt tgctatttgc acttgagaaa gagacttttc ctgcgtcgat
     1141 ggttagagaa cagtgtgcag acacttttca gctacctaga attacaattg gatatccccg
     1201 cctcccaatc cacccaccca gggaaaaaga agggctcgcc gaaaatcaaa gttatctcca
     1261 ggctcgcgca tcccaccgag cggttgactt ctctccacca cttttcattt taaccctcgg
     1321 ggtacgggat tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga
     1381 acattttcag gaggaccctt ggagggtacc gagctcagaa aaaatgactg ctccaaagaa
     1441 gaagcgtaag gtaccggtag aaaaaatgag taaaggagaa gaacttttca ctggagttgt
     1501 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
     1561 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
     1621 actacctgtt ccatgggtaa gtttaaacat atatatacta actaaccctg attatttaaa
     1681 ttttcagcca acacttgtca ctactttctg ttatggtgtt caatgcttct cgagataccc
     1741 agatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     1801 aagaactata tttttcaaag atgacgggaa ctacaagaca cgtaagttta aacagttcgg
     1861 tactaactaa ccatacatat ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac
     1921 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
     1981 acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa
     2041 gaatggaatc aaagttgtaa gtttaaacat gattttacta actaactaat ctgatttaaa
     2101 ttttcagaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca
     2161 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     2221 gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct
     2281 tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca aatagcattc
     2341 gtagaattgt aagtttaaac tttctcatac taactaacgc gctctattta aattttcaga
     2401 attccaactg agcgccggtc gctaccatta ccaacttgtc tggtgtcaaa aataataggg
     2461 gccgctgtca tcagagtaag tttaaactga gttctactaa ctaacgagta atatttaaat
     2521 tttcagcatc tcgcgcccgt gcctctgact tctaagtcca attactcttc aacatcccta
     2581 catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa aacttctctt
     2641 aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg tgtagattca
     2701 aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc ccccattaat
     2761 aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt tttttacttc
     2821 tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt atcatatgtt
     2881 acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc tcatgacgtc
     2941 aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa tcaagtgaaa
     3001 gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg tttgtcaaga
     3061 tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa
     3121 gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt gataatttga
     3181 aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc ccaatatacc
     3241 aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg
     3301 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     3361 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     3421 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     3481 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     3541 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3601 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3661 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3721 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3781 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3841 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3901 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3961 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4021 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4081 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4141 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4201 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4261 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4321 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4381 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4441 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4501 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4561 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4621 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4681 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4741 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4801 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4861 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4921 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4981 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5041 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5101 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     5161 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5221 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5281 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5341 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5401 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5461 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5521 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5581 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5641 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5701 ggaattgtga gcggataaca atttcacaca ggaaacagct
//
