LOCUS       sequence-275-exp        6800 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 6800)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 6800)
  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: L4663
FEATURES             Location/Qualifiers
     source          1..6800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2493..2513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2556..2584)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3332..3352
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4374..4396)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4496..4515
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4593..4611)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4679..4783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4784..4852,4853..5644)
                     /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(5002..5021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5815..6403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6304..6323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6557..6574
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6691..6712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6727..6732,6733..6750,6751..6757)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6765..6781
                     /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     6770..6792
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6789..6800,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6789..6800,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgggctgc aggtcggcta taataagttc ttgaataaaa
       61 taattttccc gacaaaacat gagtatttct ttcgaaaata aaagtgcagg ctaattagag
      121 attattctgt aattaactgc ataatttgtc acgtgccata gttttacatt ccactacgtc
      181 atagttctta aaatactaat ctcctgaaaa tagaagtagg tgaagaaagt ttaattatca
      241 gttctaaaat gacaattgat ctttggaata tgttctgaaa ctaccgatca ttgaacagat
      301 gctatttgaa tgatatagaa ttgtatattt gcaatttctg aaacgcgttc ttaaaggcac
      361 acagattaat tcaaaagggt ctggccgcaa aaaggtttat ggtggccgat tttgagtttt
      421 gtgtgtgatt gctttttcac aatcagtgtt ttcaggatta tgtgatgaac tagatcttca
      481 agtttcgtta catttcatat gttttcggaa ctcacgaagt acatattggg tattgtgctc
      541 aaaaaattca gcaatcagct tcgctccgct gactttagaa cccaaaaaaa tagtatggcc
      601 aaactgactg tgttacgatc atttcaattt ttcaatacat atttaagatt tctaagagta
      661 agaaggtcaa aaactgttct ggaatacata tatatttttc aggttacaat tagtcaaaaa
      721 gtgcactgaa atatacgttt taatttcacg aataacccaa ttagttcaat gtatttttgg
      781 tcaaccaacg ttaaagtttg gcttccaacc aattatcatt tctgatcaac cacaatgttt
      841 tttctttatc tgcaagttaa ttttttattt ttatccagat gtttggcata tttttcaatt
      901 cttcactagc gcccacttct tgcacttccg gcgccctgaa tctaatgcat ctgttgcaag
      961 aattgaaaga ccaatcaaca cattgttttc ttcacgagat actgaagaaa atgaataaaa
     1021 acagagaaaa agagccatgt gattagtgac aactgttgct aacagatacc atagcttgga
     1081 cttggtacgt gatggcaacg tatgggtcaa caaaaatgat tgcagagggg gtgcaaaaca
     1141 gtcaagtcga gaaaatatga aaaacagaaa acaaagaaca gaaaaatggg tttgagagtc
     1201 agtataattt ataaaagaaa aattgtacat agaaattaac catttttgta gaagaagtta
     1261 tttttcaagc atcgttaaaa attattcaaa gcaccttatt tcatatttaa ttttaaacat
     1321 ggttaaatga acaacacggt gcgcaatcag gaaaacttga aatctgaaac tgttgttgtg
     1381 atcttcttcg caactgttca gatagcacta gtgtaatgtt aagagtgcgc gaatataatg
     1441 gaatataatg gatcacacct cctgccatca ggtaaacgtc tctgttatca catatttcca
     1501 actattaaat ttttaccttt tacagtttta catttttttg aaaaaagtaa ctttttgtct
     1561 tcaaaatccc tgacgaaaat atcaaatatt ttaatcgaga ctgcagagga accgattgat
     1621 gatttggaaa atccagcttt acctgtgtaa gaactgaaaa gtttcataac cctagggtat
     1681 tcccagttac attccccact ggctaacaat agcacccagt ttttcatcac ctttcttcaa
     1741 atttctcggc gatttgttaa aaacaaaatt tgtgtccctt ctctgatatc tctatgtctc
     1801 taaacacaag ttcatcggaa aacgaaggag ggtaggtgtt ggttgggctc ccgaagtgaa
     1861 aatagaagag caagaataga atattagaga gagagtgcag agagggcggg atagctcccg
     1921 ggattccgtt ttcttcttct ttatcttcaa cgatgatgtg tgtgcgtgtt gtatagattc
     1981 tgttgctccc ccacaactcg ctccgaaggc tcaatacaat tcaattgata ttggaggaga
     2041 gcctaccgga gtgggaggat aagaagaaac ataagaagaa gaagaagaag aagcatgctt
     2101 ctggtttttg atgctatgaa aacggcacaa aaagatgatt gaggtccctt ttcaatacct
     2161 tctctcatct ttcaaatccc attgaaacct aaaacttctc accacgcttt accattgttc
     2221 tccaaaaact tatagcaatg tctataactt ttttatctct gaaaagcagt gttccatttt
     2281 tctttttcct attttatttc aattgtttct cacatttcgt ttggattctt tgcttgtcaa
     2341 ccagcttctt cttccacttt taccgtctaa ttttcagggc agggagccat caaacccacg
     2401 accactagat ccatctagag gatccccggg attggccaaa ggacccaaag gaggaccctt
     2461 ggagggtacc gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag gtaccggtag
     2521 aaaaaatgag taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
     2581 atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat
     2641 acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt ccatgggtaa
     2701 gtttaaacat atatatacta actaaccctg attatttaaa ttttcagcca acacttgtca
     2761 ctactttctc atggggcgtt caatgcttct cgagataccc agatcatatg aaacagcatg
     2821 actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag
     2881 atgacgggaa ctacaagaca cgtaagttta aacagttcgg tactaactaa ccatacatat
     2941 ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt
     3001 taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact
     3061 atatttcaca caacgtatac atcactgccg acaaacaaaa gaatgggatc aaagcggtaa
     3121 gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagaac ttcaaaatta
     3181 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     3241 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     3301 cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg
     3361 ggattacaca tggcatggat gaactataca aatagcattc gtagaattgt aagtttaaac
     3421 tttctcatac taactaacgc gctctattta aattttcaga attccaactg agcgccggtc
     3481 gctaccatta ccaacttgtc tggtgtcaaa aataataggg gccgctgtca tcagagtaag
     3541 tttaaactga gttctactaa ctaacgagta atatttaaat tttcagcatc tcgcgcccgt
     3601 gcctctgact tctaagtcca attactcttc aacatcccta catgctcttt ctccctgtgc
     3661 tcccaccccc tatttttgtt attatcaaaa aacttctctt aatttctttg ttttttagct
     3721 tcttttaagt cacctctaac aatgaaattg tgtagattca aaaatagaat taattcgtaa
     3781 taaaaagtcg aaaaaaattg tgctccctcc ccccattaat aataattcta tcccaaaatc
     3841 tacacaatgt tctgtgtaca cttcttatgt tttttacttc tgataaattt ttttgaaaca
     3901 tcatagaaaa aaccgcacac aaaatacctt atcatatgtt acgtttcagt ttatgaccgc
     3961 aatttttatt tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa
     4021 aagttttgga gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc
     4081 tgcttttgct ttttggggtt tcccctattg tttgtcaaga tttcgaggac ggcgtttttc
     4141 ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa gatcggaaga aggtttgggt
     4201 ttgaggctca gtggaaggtg agtagaagtt gataatttga aagtggagta gtgtctatgg
     4261 ggtttttgcc ttaaatgaca gaatacattc ccaatatacc aaacataact gtttcctact
     4321 agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4381 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4441 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     4501 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     4561 aggagaaaat accgcatcag gcggccttaa gggcctcgtg atacgcctat ttttataggt
     4621 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4681 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4741 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4801 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4861 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4921 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4981 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5041 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5101 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5161 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5221 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5281 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5341 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5401 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5461 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5521 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5581 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5641 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5701 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5761 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5821 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5881 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5941 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6001 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6061 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6121 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6181 cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa
     6241 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6301 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6361 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6421 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6481 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6541 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6601 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6661 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6721 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6781 atttcacaca ggaaacagct
//
