LOCUS       sequence-177-exp        6914 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 6914)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 6914)
  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: L3602
FEATURES             Location/Qualifiers
     source          1..6914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2536..2564)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3312..3332
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4354..4376)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4476..4495
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4573..4591)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4659..4763
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4764..4832,4833..5624)
                     /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(4982..5001)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5795..6383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6284..6303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6537..6554
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6671..6692
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6707..6712,6713..6730,6731..6737)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6745..6761
                     /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     6750..6772
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6769..6785
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6769..6785
                     /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 gaggatcccc gggattggcc aaaggaccca aaggtatgtt tcgaatgata ctaacataac
     2461 atagaacatt ttcaggagga cccttggagg gtaccggtag aaaaaatgag taaaggagaa
     2521 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     2581 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
     2641 tttatttgca ctactggaaa actacctgtt ccatgggtaa gtttaaacat atatatacta
     2701 actaaccctg attatttaaa ttttcagcca acacttgtca ctactttctc acacggcgta
     2761 caatgcttct cgagataccc agatcatatg aaacagcatg actttttcaa gagtgccatg
     2821 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
     2881 cgtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc aggtgctgaa
     2941 gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa
     3001 gaagatggaa acattcttgg acacaaattg gaatacaact atatttcaca caacgtatac
     3061 atcgcagcag acaaacaaaa gaatggaatc aaagttgtaa gtttaaacat gattttacta
     3121 actaactaat ctgatttaaa ttttcagaac ttcaaaatta gacacaacat tgaagatgga
     3181 agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     3241 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag
     3301 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat
     3361 gaactataca aatagcattc gtagaattgt aagtttaaac tttctcatac taactaacgc
     3421 gctctattta aattttcaga attccaactg agcgccggtc gctaccatta ccaacttgtc
     3481 tggtgtcaaa aataataggg gccgctgtca tcagagtaag tttaaactga gttctactaa
     3541 ctaacgagta atatttaaat tttcagcatc tcgcgcccgt gcctctgact tctaagtcca
     3601 attactcttc aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt
     3661 attatcaaaa aacttctctt aatttctttg ttttttagct tcttttaagt cacctctaac
     3721 aatgaaattg tgtagattca aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg
     3781 tgctccctcc ccccattaat aataattcta tcccaaaatc tacacaatgt tctgtgtaca
     3841 cttcttatgt tttttacttc tgataaattt ttttgaaaca tcatagaaaa aaccgcacac
     3901 aaaatacctt atcatatgtt acgtttcagt ttatgaccgc aatttttatt tcttcgcacg
     3961 tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa aagttttgga gtatttttgg
     4021 aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct ttttggggtt
     4081 tcccctattg tttgtcaaga tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt
     4141 gatgagcacg atgcaagaaa gatcggaaga aggtttgggt ttgaggctca gtggaaggtg
     4201 agtagaagtt gataatttga aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca
     4261 gaatacattc ccaatatacc aaacataact gtttcctact agtcggccgt acgggccctt
     4321 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4381 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4441 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     4501 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4561 gcggccttaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4621 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4681 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4741 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4801 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4861 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4921 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4981 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5041 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5101 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5161 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5221 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5281 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5341 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5401 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5461 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5521 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5581 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5641 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5701 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5941 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6181 gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6481 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6541 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     6601 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     6661 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     6721 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     6781 atgaccatga ttacgccaag ctgtaagttt aaacatgatc ttactaacta actattctca
     6841 tttaaatttt cagagcttaa aaatggctga aatcactcac aacgatggat acgctaacaa
     6901 cttggaaatg aaat
//
