LOCUS       sequence-146-exp        3616 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 3616)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 3616)
  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: L3469
FEATURES             Location/Qualifiers
     source          1..3616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(113..141)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     889..909
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1005..1022)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1005..1021)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1014..1036)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1230..1249)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1349..1371
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1409..1427)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1495..1599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1600..1668,1669..2460)
                     /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(1818..1837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2631..3219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3120..3139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3373..3390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3507..3528
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3543..3548,3549..3566,3567..3573)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3581..3597
                     /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     3586..3608
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3605..3616,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3605..3616,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag ctttcacgcg tcctggtacc gctgcagcgc ttaagttggg
       61 aattcaagta ccggtagaaa aaatgagtaa aggagaagaa cttttcactg gagttgtccc
      121 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
      181 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
      241 acctgttcca tgggtaagtt taaacatata tatactaact aaccctgatt atttaaattt
      301 tcagccaaca cttgtcacta ctctcactta tggtgttcaa tgcttctcga gatacccaga
      361 tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag
      421 aactatattt ttcaaagatg acgggaacta caagacacgt aagtttaaac agttcggtac
      481 taactaacca tacatattta aattttcagg tgctgaagtc aagtttgaag gtgataccct
      541 tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca
      601 caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa
      661 tggaatcaaa gttgtaagtt taaacatgat tttactaact aactaatctg atttaaattt
      721 tcagaacttc aaaattagac acaacattga agatggaagc gttcaactag cagaccatta
      781 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
      841 cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga
      901 gtttgtaaca gctgctggga ttacacatgg catggacgaa ctatacaaaa gcccacaagc
      961 tttcacgcgt cctggtaccg ctgcagcgct taagttggga attcactggc cgtcgtttta
     1021 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     1081 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     1141 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     1201 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     1261 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     1321 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     1381 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     1441 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1561 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1621 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1681 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1741 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1801 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1861 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1921 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1981 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2041 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2101 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2161 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2221 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2281 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2341 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2401 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2461 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2521 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2581 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2641 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2701 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2761 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2821 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2881 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2941 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3001 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3061 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3241 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3301 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3361 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3421 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3481 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     3541 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     3601 cacacaggaa acagct
//
