LOCUS       sequence-241-exp        5094 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 5094)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 5094)
  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: L4053
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             527..547
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             569..589
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             611..631
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             653..673
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(716..744)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1492..1512
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2534..2556)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2656..2675
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2753..2771)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2839..2943
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2944..3012,3013..3804)
                     /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(3162..3181)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3975..4563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4464..4483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4717..4734
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4851..4872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4887..4892,4893..4910,4911..4917)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4925..4941
                     /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     4930..4952
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4949..4965
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4949..4965
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagaggat ccccggggat tggccaaagg
       61 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
      121 gcttggctag caaaaatgca taaggttttg ctggcactgt tctttatctt tctggcacca
      181 gcatccgcac tggcagtctc cgaaccggcc tgcaggatcg attttttgca aattacgagc
      241 gttgtagggg gcggagcgat aggtcctata ggttttggta tatcatcatt cattcattca
      301 ttggtacatt catttaccca ccttcctctt tctgagcttc tctggagttc tgtgcttcct
      361 ttttccctta tctttatact gtaattttta actttcaggc attgattgga tccccgggat
      421 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      481 gaggaccctt gcttggaggg taccgagctc agaaaaaatg actgctccaa agaagaagcg
      541 taaggtaccg gcagaaaaaa tgactgctcc aaagaagaag cgtaaggtac cggcagaaaa
      601 aatgactgct ccaaagaaga agcgtaaggt accggcagaa aaaatgactg ctccaaagaa
      661 gaagcgtaag gtaccggtag aaaaaatgag taaaggagaa gaacttttca ctggagttgt
      721 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
      781 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
      841 actacctgtt ccatgggtaa gtttaaacat atatatacta actaaccctg attatttaaa
      901 ttttcagcca acacttgtca ctactttctg ttatggtgtt caatgcttct cgagataccc
      961 agatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     1021 aagaactata tttttcaaag atgacgggaa ctacaagaca cgtaagttta aacagttcgg
     1081 tactaactaa ccatacatat ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac
     1141 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
     1201 acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa
     1261 gaatggaatc aaagttgtaa gtttaaacat gattttacta actaactaat ctgatttaaa
     1321 ttttcagaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca
     1381 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
     1441 gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct
     1501 tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca aatagcattc
     1561 gtagaattgt aagtttaaac tttctcatac taactaacgc gctctattta aattttcaga
     1621 attccaactg agcgccggtc gctaccatta ccaacttgtc tggtgtcaaa aataataggg
     1681 gccgctgtca tcagagtaag tttaaactga gttctactaa ctaacgagta atatttaaat
     1741 tttcagcatc tcgcgcccgt gcctctgact tctaagtcca attactcttc aacatcccta
     1801 catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa aacttctctt
     1861 aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg tgtagattca
     1921 aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc ccccattaat
     1981 aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt tttttacttc
     2041 tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt atcatatgtt
     2101 acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc tcatgacgtc
     2161 aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa tcaagtgaaa
     2221 gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg tttgtcaaga
     2281 tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa
     2341 gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt gataatttga
     2401 aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc ccaatatacc
     2461 aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg
     2521 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2581 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2641 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2701 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     2761 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     2821 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2881 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2941 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3001 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3061 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3121 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3181 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3241 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3301 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3361 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3421 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3481 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3541 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3601 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3661 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3721 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3781 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3841 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3901 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3961 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4021 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4081 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4141 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4201 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4261 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4321 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     4381 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4441 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4501 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4561 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4621 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4681 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4741 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     4801 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     4861 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     4921 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     4981 ctgtaagttt aaacatgatc ttactaacta actattctca tttaaatttt cagagcttaa
     5041 aaatggctga aatcactcac aacgatggat acgctaacaa cttggaaatg aaat
//
