LOCUS       sequence-296-exp        3410 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 3410)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 3410)
  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: L754
FEATURES             Location/Qualifiers
     source          1..3410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(984..1006)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1106..1125
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1203..1221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1289..1393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1394..1462,1463..2254)
                     /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(1612..1631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2425..3013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2914..2933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3167..3184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3301..3322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3337..3342,3343..3360,3361..3367)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3375..3391
                     /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     3380..3402
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3399..3410,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3399..3410,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggat
       61 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      121 gaggaccctt ggctagcgtc gacggtacca tggtattgat atctgagctc cgcatcggcc
      181 gctgtcatca gatcgccatc tcgcgcccgt gcctctgact tctaagtcca attactcttc
      241 aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa
      301 aacttctctt aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg
      361 tgtagattca aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc
      421 ccccattaat aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt
      481 tttttacttc tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt
      541 atcatatgtt acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc
      601 tcatgacgtc aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa
      661 tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg
      721 tttgtcaaga tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg
      781 atgcaagaaa gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt
      841 gataatttga aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc
      901 ccaatatacc aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg
      961 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     1021 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     1081 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     1141 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa
     1201 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1261 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1321 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1381 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1441 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1501 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1561 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1621 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1681 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1741 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1801 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1861 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1921 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1981 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2041 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2101 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2161 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2221 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg
     2821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3121 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3181 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3241 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3301 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     3361 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
//
