LOCUS       sequence-56920-e        3027 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 3027)
  AUTHORS   Hirani N, Westenberg M, Gami MS, Davis P, Hope IA, Dolphin CT
  TITLE     A simplified counter-selection recombineering protocol for creating 
            fluorescent protein reporter constructs directly from C. elegans 
            fosmid genomic clones.
  JOURNAL   BMC Biotechnol. 2013 Jan 3;13(1):1.
  PUBMED    23281894
REFERENCE   2  (bases 1 to 3027)
  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"; journalName: "BMC 
            Biotechnol."; date: "2013-01-3"; volume: "13(1)"; pages: "1"
COMMENT     Sequence Label: pNH009
FEATURES             Location/Qualifiers
     source          1..3027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(856..1647,1648..1716))
                     /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     1479..1498
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1717..1821)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1964..2008
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2024..2047
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2063..2086
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(join(2856..2862,2863..2880,2881..2886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2901..2922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgaatt aattcttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
      901 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      961 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1021 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1081 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1141 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1201 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1261 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1321 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1381 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1441 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1501 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1561 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1621 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1681 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1741 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1801 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
     1861 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1921 cccctcagcg cggccgctag cggatcctta aaaaaccatg gttaaagaaa cagcagcagc
     1981 gaaatttgaa agacaacata tggatagtgg aggcggtgga agtttggaag ttcttttcca
     2041 aggaccgggc ggaggaggct cttggtcgca tccacagttc gaaaaagcca tggccgcctc
     2101 aaaaggagaa gaaacgacaa tgggtgttat caagccagac atgaagatca agcttaaaat
     2161 ggagggtaat gttaatggtc acgccttcgt cattgaggga gagggtgaag gaaagccata
     2221 cgatggaact aatactatta acttggaagt gaaggagggt gccccacttc cattctctta
     2281 tgatatcctt actacagcct tcgcatatgg aaatcgagca ttcacaaaat acccagacga
     2341 cattccaaat tacttcaagc agtcatttcc agagggatat tcttgggaga gaacaatgac
     2401 attcgaggac aagggtatag ttaaagtgaa atctgatatt agtatggaag aagattcatt
     2461 catctatgag atacacctga agggcgagaa ctttcctccg aatggaccag tgatgcagaa
     2521 aaagacaaca ggatgggacg caagtactga acgtatgtat gtcagagatg gagttttgaa
     2581 aggtgacgtg aagcataaac tgttgttgga aggaggtgga catcatcgag tggactttaa
     2641 gacaatctac agagctaaga aagctgtgaa attgccagac tatcactttg tggatcaccg
     2701 aattgaaatt ttgaaccatg ataaagatta caataaagtg actgtgtatg agtcagcagt
     2761 agctagaaac tcaactgatg gtatggacga gctctacgta agcgcttaat aggtaccgcg
     2821 gccgcgctga ggggctggcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2881 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2941 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3001 gggagaggcg gtttgcgtat tgggcgc
//
