LOCUS       sequence-247-exp        6898 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 6898)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 6898)
  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: L4063
FEATURES             Location/Qualifiers
     source          1..6898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2520..2548)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3296..3316
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4338..4360)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4460..4479
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4557..4575)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4643..4747
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4748..4816,4817..5608)
                     /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(4966..4985)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5779..6367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6268..6287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6521..6538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6655..6676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6691..6696,6697..6714,6715..6721)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6729..6745
                     /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     6734..6756
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6753..6769
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6753..6769
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcaggtcgg ctataataag ttcttgaata aaataatttt cccgacaaaa
       61 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      121 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      181 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      241 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      301 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      361 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      421 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      481 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      541 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      601 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      661 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      721 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      781 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      841 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
      901 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
      961 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1021 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1081 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1141 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1201 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1261 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1321 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1381 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1441 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1501 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1561 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1621 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1681 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1741 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1801 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1861 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     1921 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     1981 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2041 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2101 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2161 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2221 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2281 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2341 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2401 gaccgcggtt ttggcgcgga tcctatcgat tcgcggccgc tgtacacccg ggtgccctat
     2461 agtgagtcgt attggtaccg gtagaaaaaa tgagtaaagg agaagaactt ttcactggag
     2521 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     2581 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     2641 gaaaactacc tgttccatgg gtaagtttaa acatatatat actaactaac cctgattatt
     2701 taaattttca gccaacactt gtcactactt tctgctgggg cgtacaatgc ttctcgagat
     2761 acccagatca tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac
     2821 aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtaag tttaaacagt
     2881 tcggtactaa ctaaccatac atatttaaat tttcaggtgc tgaagtcaag tttgaaggtg
     2941 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
     3001 ttggacacaa attggaatac aactatattt cacacaacgt atacatcact gccgacaaac
     3061 aaaagaatgg gatcaaagcg gtaagtttaa acatgatttt actaactaac taatctgatt
     3121 taaattttca gaacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag
     3181 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
     3241 acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc
     3301 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatagc
     3361 attcgtagaa ttgtaagttt aaactttctc atactaacta acgcgctcta tttaaatttt
     3421 cagaattcca actgagcgcc ggtcgctacc attaccaact tgtctggtgt caaaaataat
     3481 aggggccgct gtcatcagag taagtttaaa ctgagttcta ctaactaacg agtaatattt
     3541 aaattttcag catctcgcgc ccgtgcctct gacttctaag tccaattact cttcaacatc
     3601 cctacatgct ctttctccct gtgctcccac cccctatttt tgttattatc aaaaaacttc
     3661 tcttaatttc tttgtttttt agcttctttt aagtcacctc taacaatgaa attgtgtaga
     3721 ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat
     3781 taataataat tctatcccaa aatctacaca atgttctgtg tacacttctt atgtttttta
     3841 cttctgataa atttttttga aacatcatag aaaaaaccgc acacaaaata ccttatcata
     3901 tgttacgttt cagtttatga ccgcaatttt tatttcttcg cacgtctggg cctctcatga
     3961 cgtcaaatca tgctcatcgt gaaaaagttt tggagtattt ttggaatttt tcaatcaagt
     4021 gaaagtttat gaaattaatt ttcctgcttt tgctttttgg ggtttcccct attgtttgtc
     4081 aagatttcga ggacggcgtt tttcttgcta aaatcacaag tattgatgag cacgatgcaa
     4141 gaaagatcgg aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga agttgataat
     4201 ttgaaagtgg agtagtgtct atggggtttt tgccttaaat gacagaatac attcccaata
     4261 taccaaacat aactgtttcc tactagtcgg ccgtacgggc cctttcgtct cgcgcgtttc
     4321 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     4381 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     4441 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     4501 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct
     4561 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4621 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4681 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4741 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4801 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4861 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4921 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4981 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5041 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5101 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5161 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5221 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5281 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5341 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5401 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5461 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5521 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5581 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5641 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5701 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5761 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5821 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5881 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     5941 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6001 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6061 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6121 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     6181 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6241 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6301 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6361 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6421 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6481 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6541 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6601 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6661 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     6721 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     6781 caagctgtaa gtttaaacat gatcttacta actaactatt ctcatttaaa ttttcagagc
     6841 ttaaaaatgg ctgaaatcac tcacaacgat ggatacgcta acaacttgga aatgaaat
//
