LOCUS       sequence-285-exp        4650 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 4650)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4650)
  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: L4752
FEATURES             Location/Qualifiers
     source          1..4650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(518..546)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1294..1314
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        2395..2499
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2500..2568,2569..3360)
                     /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(2718..2737)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3531..4119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4020..4039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4273..4290
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4407..4428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4443..4448,4449..4466,4467..4473)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4481..4497
                     /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     4486..4508
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4505..4521
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4505..4521
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc cccgggattg
       61 gccaaaggac ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga
      121 ggacccttgg agggtaccta atttgatcga taatatgtac atattttgtt attcgttccg
      181 gtatcgtacc tccacctacc ataaatccca taccctacat ctttatcgat tttttgcaaa
      241 ttacgagcgt tgtagggggc ggagcgatag gtcctatagg ttttggtata tcatcattca
      301 ttcattcatt ggtacattca tttacccacc ttcctctttc tgagcttctc tggagttctg
      361 tgcttccttt ttcccttatc tttatactgt aatttttaac tttcaggcat tgattggatc
      421 ctatcgattc gcggccgctg tacacccggg tgccctatag tgagtcgtat tggtaccggt
      481 agaaaaaatg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
      541 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
      601 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatgggt
      661 aagtttaaac atatatatac taactaaccc tgattattta aattttcagc caacacttgt
      721 cactactttc tcatggggcg ttcaatgctt ctcgagatac ccagatcata tgaaacagca
      781 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa
      841 agatgacggg aactacaaga cacgtaagtt taaacagttc ggtactaact aaccatacat
      901 atttaaattt tcaggtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga
      961 gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa
     1021 ctatatttca cacaacgtat acatcactgc cgacaaacaa aagaatggga tcaaagcggt
     1081 aagtttaaac atgattttac taactaacta atctgattta aattttcaga acttcaaaat
     1141 tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc
     1201 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct
     1261 ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc
     1321 tgggattaca catggcatgg atgaactata caaatagcat tcgtagaatt gtaagtttaa
     1381 actttctcat actaactaac gcgctctatt taaattttca gaattccaac tgagcgccgg
     1441 tcgctaccat taccaacttg tctggtgtca aaaataatag gggccgctgt catcagagta
     1501 agtttaaact gagttctact aactaacgag taatatttaa attttcagca tctcgcgccc
     1561 gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct ttctccctgt
     1621 gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt tgttttttag
     1681 cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga attaattcgt
     1741 aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc tatcccaaaa
     1801 tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat ttttttgaaa
     1861 catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca gtttatgacc
     1921 gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg ctcatcgtga
     1981 aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga aattaatttt
     2041 cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg acggcgtttt
     2101 tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg
     2161 gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag tagtgtctat
     2221 ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa ctgtttccta
     2281 ctagtaaaaa ataagcaaag aaattaggcg aacgatgaga attgtcctcg cttgggccca
     2341 aaaggccttt tttttttggc gccgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2401 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2461 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2521 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2581 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2641 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2701 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2761 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2821 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2881 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2941 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3001 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3061 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3121 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3181 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3241 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3301 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3361 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3421 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3481 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3541 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3601 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3661 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3721 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3781 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3841 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3901 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3961 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4021 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4081 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4141 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4201 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4261 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4321 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4381 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4441 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4501 cacacaggaa acagctatga ccatgattac gccaagctgt aagtttaaac atgatcttac
     4561 taactaacta ttctcattta aattttcaga gcttaaaaat ggctgaaatc actcacaacg
     4621 atggatacgc taacaacttg gaaatgaaat
//
