LOCUS       sequence-180-exp        4810 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 4810)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 4810)
  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: L3605
FEATURES             Location/Qualifiers
     source          1..4810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             369..389
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(432..460)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1208..1228
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2250..2272)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2372..2391
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2469..2487)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2555..2659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2660..2728,2729..3520)
                     /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(2878..2897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3691..4279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4180..4199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4433..4450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4567..4588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4603..4608,4609..4626,4627..4633)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4641..4657
                     /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     4646..4668
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4665..4681
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4665..4681
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccaa ttcttgagac ttccactatc
       61 ttctctggca ccctccacta cgcgtaggcg tctctcgctt cgtgtattcc cgggaagccg
      121 ggttcccgtc tctcccgccg ctgccgctgc cgcacacagc tttacacctc gtagaatccc
      181 caaagagggg cgtggcttgc gggtgccaac atcctcctgc cgaggaagaa gcaggcactc
      241 atcactcgca tcatcaacct cgggattggc caaaggaccc aaaggtatgt ttcgaatgat
      301 actaacataa catagaacat tttcaggagg acccttggag ggtaccgagc tcagaaaaaa
      361 tgactgctcc aaagaagaag cgtaaggtac cggtagaaaa aatgagtaaa ggagaagaac
      421 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
      481 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
      541 tttgcactac tggaaaacta cctgttccat gggtaagttt aaacatatat atactaacta
      601 accctgatta tttaaatttt cagccaacac ttgtcactac tttcacttat ggtgttcaat
      661 gcttctcgag atacccagat catatgaaac agcatgactt tttcaagagt gccatgcccg
      721 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgta
      781 agtttaaaca gttcggtact aactaaccat acatatttaa attttcaggt gctgaagtca
      841 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
      901 atggaaacat tcttggacac aaattggaat acaactatat ttcacacaac gtatacatca
      961 ctgccgacaa acaaaagaat gggatcaaag cggtaagttt aaacatgatt ttactaacta
     1021 actaatctga tttaaatttt cagaacttca aaattagaca caacattgaa gatggaagcg
     1081 ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac
     1141 cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag
     1201 accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac
     1261 tatacaaata gcattcgtag aattgtaagt ttaaactttc tcatactaac taacgcgctc
     1321 tatttaaatt ttcagaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt
     1381 gtcaaaaata ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa
     1441 cgagtaatat ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta
     1501 ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta
     1561 tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg
     1621 aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct
     1681 ccctcccccc attaataata attctatccc aaaatctaca caatgttctg tgtacacttc
     1741 ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa
     1801 taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg
     1861 ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt
     1921 tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc
     1981 ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg
     2041 agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta
     2101 gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat
     2161 acattcccaa tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt
     2221 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     2281 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     2341 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     2401 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg
     2461 ccttaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2521 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2581 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2641 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2701 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2761 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2821 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2881 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2941 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3001 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3061 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3121 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3181 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3241 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3301 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3361 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3421 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3481 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3541 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3601 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3661 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3721 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3781 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3841 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3901 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3961 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4021 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4081 gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4141 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4201 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4261 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4321 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4381 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4441 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     4501 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     4561 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     4621 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     4681 ccatgattac gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta
     4741 aattttcaga gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg
     4801 gaaatgaaat
//
