LOCUS       sequence-200-exp        3777 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 3777)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 3777)
  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: L3781
FEATURES             Location/Qualifiers
     source          1..3777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(148..176)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     924..944
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        1522..1626
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1627..1695,1696..2487)
                     /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(1845..1864)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2658..3246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3147..3166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3400..3417
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3534..3555
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3570..3575,3576..3593,3594..3600)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3608..3624
                     /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     3613..3635
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3632..3648
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3632..3648
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc ctatcgattc
       61 gcggccgctg tacacccggg tgccctatag tgagtcgtat tggtaccggt agaaaaaatg
      121 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
      181 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa
      241 cttaccctta aatttatttg cactactgga aaactacctg ttccatgggt aagtttaaac
      301 atatatatac taactaaccc tgattattta aattttcagc caacacttgt cactactttc
      361 tgttatggtg ttcaatgctt ctcgagatac ccagatcata tgaaacagca tgactttttc
      421 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
      481 aactacaaga cacgtaagtt taaacagttc ggtactaact aaccatacat atttaaattt
      541 tcaggtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
      601 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca
      661 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttgt aagtttaaac
      721 atgattttac taactaacta atctgattta aattttcaga acttcaaaat tagacacaac
      781 attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat
      841 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
      901 cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca
      961 catggcatgg acgaactata caaaagccca caagctttgg aattcgctag ccggccatac
     1021 aagtaatccg gatgatcgac gccaacgtcg ttgaattttc aaattttaaa tactgaatat
     1081 ttgttttttt tcctattatt tatttattct ctttgtgttt tttttcttgc tttctaaaaa
     1141 attaattcaa tccaaatcta aacatttttt tttctctttc cgtctcccaa ttcgtattcc
     1201 gctcctctca tctgaacaca atgtgcaagt ttatttatct tctcgctttc atttcattag
     1261 gacgtggggg gaattggtgg aagggggaaa cacacaaaag gatgatggaa atgaaataag
     1321 gacacacaat atgcaacaac attcaattca gaaatatgga ggaaggttta aaagaaaaca
     1381 taaaaatata tagaggagga aggaaaacta gtaaaaaata agcaaagaaa ttaggcgaac
     1441 gatgagaatt gtcctcgctt gggcccaaaa ggcctttttt ttttggcgcc gacgtcaggt
     1501 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1561 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1621 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1681 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1741 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1801 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1861 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1921 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1981 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2041 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2101 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2161 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2221 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2281 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2341 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2401 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2461 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2521 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2581 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2641 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2701 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2761 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2821 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2881 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2941 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3001 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     3061 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3121 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3181 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3241 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3301 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3361 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3421 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3481 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     3541 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     3601 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     3661 aagctgtaag tttaaacatg atcttactaa ctaactattc tcatttaaat tttcagagct
     3721 taaaaatggc tgaaatcact cacaacgatg gatacgctaa caacttggaa atgaaat
//
