LOCUS       sequence-216-exp        4568 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 4568)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4568)
  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: L2528
FEATURES             Location/Qualifiers
     source          1..4568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2008..2030)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2130..2149
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2227..2245)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2313..2417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2418..2486,2487..3278)
                     /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(2636..2655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3449..4037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3938..3957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4191..4208
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4325..4346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4361..4366,4367..4384,4385..4391)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4399..4415
                     /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     4404..4426
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4423..4439
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4423..4439
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga aatgtaaacc tgtcatttct gtgtatttca
       61 gcaccgcgaa gagcaccata aacagctaca acaagagcac gaacgtcggc tccaacagaa
      121 aaagctgcaa aacgctggta ctcaccacat gctctgataa ttgccatttc cttcgatttc
      181 tgactttcaa ttgtgatatg ataacctcaa aatctccatt caaaatgaaa actcttaaat
      241 ttaagagctt tcaacacccc gcactttccc acctcatcgt ttttcccttc tatcttatca
      301 attttttgta ttttgtgtca gtttgctata tcctcatcac cttttttacc gtttaatctc
      361 tctatagtat tgttcgggtg tttcaagatg aaacgttttg tgtgtagcat ttttgaaaca
      421 cagcgcgaaa aatgcagaaa ctatcatcca caatgcatta agtgcactca ttgtgatgac
      481 tcctaacccc ctcgccccac cagttgtctc ttcacaaatt catggcacaa ttcaaaaatg
      541 gctttttgtg ttggatcttg atctcgtctc tttccttctt ctttcgtttc tctcggacca
      601 ctcattattc gaatgccatg catttgttga tgatgcgcag cgatggtcgc acacactaca
      661 acagatatga tggtggctgt gcgtggtgca gagctcatga ataagtgatg gggcaccgag
      721 atgtgactcc tcccattatt cgtcttgagt cgcatctttt gtcttggtcc gttgtcagtc
      781 gagcctcaag gcagccgttc cgccagtgat cgcctgctaa gaagttgtag gtgtagagga
      841 agaaacgtcg ggtccaaatt tcaagcaaca tccgcgagat cgtgaatccc ggggattggc
      901 caaaggaccc aaaggtatgt ttcgaatgat actaacataa catagaacat tttcaggagg
      961 acccttggct agcgtcgacg gtaccggtca gatatcactc gaggagctcc cgagatcttc
     1021 agcgctcagc attcgtagaa ttgtaagttt aaactttctc atactaacta acgcgctcta
     1081 tttaaatttt cagaattcca actgagcgcc ggtcgctacc attaccaact tgtctggtgt
     1141 caaaaataat aggggccgct gtcatcagag taagtttaaa ctgagttcta ctaactaacg
     1201 agtaatattt aaattttcag catctcgcgc ccgtgcctct gacttctaag tccaattact
     1261 cttcaacatc cctacatgct ctttctccct gtgctcccac cccctatttt tgttattatc
     1321 aaaaaacttc tcttaatttc tttgtttttt agcttctttt aagtcacctc taacaatgaa
     1381 attgtgtaga ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc
     1441 ctccccccat taataataat tctatcccaa aatctacaca atgttctgtg tacacttctt
     1501 atgtttttta cttctgataa atttttttga aacatcatag aaaaaaccgc acacaaaata
     1561 ccttatcata tgttacgttt cagtttatga ccgcaatttt tatttcttcg cacgtctggg
     1621 cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt tggagtattt ttggaatttt
     1681 tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt tgctttttgg ggtttcccct
     1741 attgtttgtc aagatttcga ggacggcgtt tttcttgcta aaatcacaag tattgatgag
     1801 cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga
     1861 agttgataat ttgaaagtgg agtagtgtct atggggtttt tgccttaaat gacagaatac
     1921 attcccaata taccaaacat aactgtttcc tactagtcgg ccgtacgggc cctttcgtct
     1981 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2041 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2101 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2161 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcggcc
     2221 ttaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2281 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2341 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2401 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2461 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2521 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2581 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2641 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2701 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2761 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2821 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2881 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2941 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3001 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3061 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3121 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3181 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3241 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3301 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3361 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3841 attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4141 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4201 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4261 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4321 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     4381 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     4441 atgattacgc caagctgtaa gtttaaacat gatcttacta actaactatt ctcatttaaa
     4501 ttttcagagc ttaaaaatgg ctgaaatcac tcacaacgat ggatacgcta acaacttgga
     4561 aatgaaat
//
