LOCUS       sequence-228-exp        5128 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 5128)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 5128)
  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: L4035
FEATURES             Location/Qualifiers
     source          1..5128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(805..833)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1581..1601
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2568..2590)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2690..2709
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2787..2805)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2873..2977
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2978..3046,3047..3838)
                     /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(3196..3215)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4009..4597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4498..4517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4751..4768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4885..4906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4921..4926,4927..4944,4945..4951)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4959..4975
                     /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     4964..4986
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4983..4999
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4983..4999
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagaggat ccccggggat tggccaaagg
       61 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
      121 gcttggctag caaaaatgca taaggttttg ctggcactgt tctttatctt tctggcacca
      181 gcatccgcac tggcagtctc cgaaccggcc tgcaggatcg attttttgca aattacgagc
      241 gttgtagggg gcggagcgat aggtcctata ggttttggta tatcatcatt cattcattca
      301 ttggtacatt catttaccca ccttcctctt tctgagcttc tctggagttc tgtgcttcct
      361 ttttccctta tctttatact gtaattttta actttcaggc attgattgga tccccgggat
      421 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      481 gaggaccctt gcttggaggg taccgagccg gcaatgccac cttcaacatc attgctgctc
      541 ctcgcagcac ttcttccatt cgctttacca gcaagcgatt ggaagactgg agaagtcact
      601 ccggataatg caacagtctg ggtcagaaaa cataaagatt gtcctccacc tgtccctgtg
      661 ctcgcaattg tgctcggagt cattgcgggt atcgtaatcc tcggaattct tctcttgttg
      721 ctctggaaat tgctcacagt acttcatgat agatccgagg taccggtaga aaaaatgagt
      781 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
      841 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
      901 acccttaaat ttatttgcac tactggaaaa ctacctgttc catgggtaag tttaaacata
      961 tatatactaa ctaaccctga ttatttaaat tttcagccaa cacttgtcac tactttctgt
     1021 tatggtgttc aatgcttctc gagataccca gatcatatga aacggcatga ctttttcaag
     1081 agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac
     1141 tacaagacac gtaagtttaa acagttcggt actaactaac catacatatt taaattttca
     1201 ggtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt
     1261 gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
     1321 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttgtaag tttaaacatg
     1381 attttactaa ctaactaatc tgatttaaat tttcagaact tcaaaattag acacaacatt
     1441 gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc
     1501 cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc
     1561 aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat
     1621 ggcatggatg aactatacaa atagcattcg tagaattcca actgagcgcc ggtcgctacc
     1681 attaccaact tgtctggtgt caaaaataat aggggccgct gtcatcagag taagtttaaa
     1741 ctgagttcta ctaactaacg agtaatattt aaattttcag catctcgcgc ccgtgcctct
     1801 gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct gtgctcccac
     1861 cccctatttt tgttattatc aaaaaacttc tcttaatttc tttgtttttt agcttctttt
     1921 aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc gtaataaaaa
     1981 gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa aatctacaca
     2041 atgttctgtg tacacttctt atgtttttta cttctgataa atttttttga aacatcatag
     2101 aaaaaaccgc acacaaaata ccttatcata tgttacgttt cagtttatga ccgcaatttt
     2161 tatttcttcg cacgtctggg cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt
     2221 tggagtattt ttggaatttt tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt
     2281 tgctttttgg ggtttcccct attgtttgtc aagatttcga ggacggcgtt tttcttgcta
     2341 aaatcacaag tattgatgag cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg
     2401 ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg agtagtgtct atggggtttt
     2461 tgccttaaat gacagaatac attcccaata taccaaacat aactgtttcc tactagtcgg
     2521 ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     2581 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     2641 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     2701 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2761 aaataccgca tcaggcggcc ttaagggcct cgtgatacgc ctatttttat aggttaatgt
     2821 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     2881 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2941 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3001 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3061 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3121 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3181 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3241 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3301 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3361 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3421 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3481 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3541 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3601 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3661 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3721 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3781 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3841 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3901 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3961 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4021 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4081 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4141 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4201 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4261 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4321 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4381 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4441 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4501 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4561 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4621 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4681 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4741 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     4801 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     4861 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     4921 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     4981 cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat gatcttacta
     5041 actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac tcacaacgat
     5101 ggatacgcta acaacttgga aatgaaat
//
