LOCUS       sequence-149-exp        4097 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 4097)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 4097)
  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: L3503
FEATURES             Location/Qualifiers
     source          1..4097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..41
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(111..127)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(230..258)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1006..1026
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1125..1141
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1211..1227)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1252..1271)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1253..1271)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1281..1298)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1281..1297)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1290..1312)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1439..1894
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1526..1545)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1736..1757
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1920..2024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2025..2093,2094..2885)
                     /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(2243..2262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3056..3644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3545..3564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3798..3815
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3932..3953
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3968..3973,3974..3991,3992..3998)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4006..4022
                     /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     4011..4033
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4030..4046
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4030..4046
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4065..4085
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4067..4085
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctccacc gcggtggcgg ccgctctaga actagtggat ccaccggttc catggctagc
       61 cacgtgacgc gtggatcccc cgggctgcag gaattcgata tcaagcttat cgataccgtc
      121 gacctcgagg gggggcccgg tacgatatac taactaacaa gaattatcta atttttcagg
      181 tccggtaccg gtagaaaaaa tgagtaaagg agaagaactt ttcactggag ttgtcccaat
      241 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
      301 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
      361 tgttccatgg gtaagtttaa acatatatat actaactaac cctgattatt taaattttca
      421 gccaacactt gtcactactc tcacttatgg tgttcaatgc ttctcgagat acccagatca
      481 tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
      541 tatatttttc aaagatgacg ggaactacaa gacacgtaag tttaaacagt tcggtactaa
      601 ctaaccatac atatttaaat tttcaggtgc tgaagtcaag tttgaaggtg atacccttgt
      661 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
      721 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
      781 aatcaaagtt gtaagtttaa acatgatttt actaactaac taatctgatt taaattttca
      841 gaacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
      901 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
      961 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     1021 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaaggta agttttaaca
     1081 tttaatcata cgaaatgaaa gagctccacc gcggtggcgg ccgctctaga actagtggat
     1141 ccaccggttc catggctagc cacgtgacgc gtggatcccc cgggctgcag gaattcgata
     1201 tcaagcttat cgataccgtc gacctcgagg gggggcccgg tacccaattc gccctatagt
     1261 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1321 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1381 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     1441 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1501 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1561 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1621 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1681 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1741 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1801 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1861 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1921 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1981 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2041 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2101 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2161 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2221 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2281 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2341 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2401 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2461 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2521 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2581 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2641 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2701 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2761 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2821 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2881 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2941 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3001 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3061 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3181 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3721 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3781 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3841 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3901 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     3961 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     4021 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     4081 aaagggaaca aaagctg
//
