LOCUS       sequence-242-exp        4713 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 4713)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4713)
  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: L4054
FEATURES             Location/Qualifiers
     source          1..4713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             146..166
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             188..208
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             230..250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             272..292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(335..363)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1111..1131
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2153..2175)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2275..2294
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2372..2390)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2458..2562
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2563..2631,2632..3423)
                     /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(2781..2800)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3594..4182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4083..4102
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4336..4353
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4470..4491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4506..4511,4512..4529,4530..4536)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4544..4560
                     /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     4549..4571
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4568..4584
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4568..4584
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gattggccaa aggacccaaa
       61 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      121 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg cagaaaaaat
      181 gactgctcca aagaagaagc gtaaggtacc ggcagaaaaa atgactgctc caaagaagaa
      241 gcgtaaggta ccggcagaaa aaatgactgc tccaaagaag aagcgtaagg taccggtaga
      301 aaaaatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
      361 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata
      421 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catgggtaag
      481 tttaaacata tatatactaa ctaaccctga ttatttaaat tttcagccaa cacttgtcac
      541 tactttctgt tatggtgttc aatgcttctc gagataccca gatcatatga aacggcatga
      601 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga
      661 tgacgggaac tacaagacac gtaagtttaa acagttcggt actaactaac catacatatt
      721 taaattttca ggtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
      781 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
      841 taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca aagttgtaag
      901 tttaaacatg attttactaa ctaactaatc tgatttaaat tttcagaact tcaaaattag
      961 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     1021 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     1081 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     1141 gattacacat ggcatggatg aactatacaa atagcattcg tagaattgta agtttaaact
     1201 ttctcatact aactaacgcg ctctatttaa attttcagaa ttccaactga gcgccggtcg
     1261 ctaccattac caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat cagagtaagt
     1321 ttaaactgag ttctactaac taacgagtaa tatttaaatt ttcagcatct cgcgcccgtg
     1381 cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc tccctgtgct
     1441 cccaccccct atttttgtta ttatcaaaaa acttctctta atttctttgt tttttagctt
     1501 cttttaagtc acctctaaca atgaaattgt gtagattcaa aaatagaatt aattcgtaat
     1561 aaaaagtcga aaaaaattgt gctccctccc cccattaata ataattctat cccaaaatct
     1621 acacaatgtt ctgtgtacac ttcttatgtt ttttacttct gataaatttt tttgaaacat
     1681 catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt tatgaccgca
     1741 atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc atcgtgaaaa
     1801 agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat taattttcct
     1861 gcttttgctt tttggggttt cccctattgt ttgtcaagat ttcgaggacg gcgtttttct
     1921 tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa ggtttgggtt
     1981 tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag tgtctatggg
     2041 gtttttgcct taaatgacag aatacattcc caatatacca aacataactg tttcctacta
     2101 gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2161 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2221 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     2281 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     2341 ggagaaaata ccgcatcagg cggccttaag ggcctcgtga tacgcctatt tttataggtt
     2401 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2461 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2521 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2581 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2641 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2701 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2761 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2821 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2881 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2941 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3001 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3061 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3121 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3181 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3241 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3301 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3361 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3421 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3481 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3541 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3601 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3661 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3721 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3781 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3841 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3901 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3961 gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag
     4021 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4081 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4141 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4201 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4261 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4321 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     4381 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     4441 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     4501 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     4561 tttcacacag gaaacagcta tgaccatgat tacgccaagc tgtaagttta aacatgatct
     4621 tactaactaa ctattctcat ttaaattttc agagcttaaa aatggctgaa atcactcaca
     4681 acgatggata cgctaacaac ttggaaatga aat
//
