LOCUS       sequence-56937-e        9267 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 9267)
  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 9267)
  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: pNH042
FEATURES             Location/Qualifiers
     source          1..9267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(240..1031,1032..1100))
                     /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     863..882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1101..1205)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(join(2258..2264,2265..2282,2283..2288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2303..2324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(2544..3203)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3118..3137
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3204..3306)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4164..4778
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      4255..4786
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      4854..5073
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             5164..5919
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    5922..6172
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             6498..7673
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             7673..8644
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    8717..9190
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
ORIGIN
        1 agaacggagt aacctcggtg tgcggttgta tgcctgctgt ggattgctgc tgtgtcctgc
       61 ttatccacaa cattttgcgc acggttatgt ggacaaaata cctggttacc caggccgtgc
      121 cggcacgtta accgggctgc atccgatgca agtgtgtcgc tgtcgacgag ctcgcgagct
      181 cgtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg aattaattct
      241 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
      301 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
      361 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
      421 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
      481 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
      541 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
      601 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
      661 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
      721 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
      781 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
      841 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
      901 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
      961 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1021 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1081 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1141 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     1201 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     1261 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcccctc agcgcggccg
     1321 ctagcggatc cttaaaaatg gtctcaaagg gtgaagaaga taacatggca attattaaag
     1381 agtttatgcg tttcaaggtg catatggagg gatctgtcaa tgggcatgag tttgaaattg
     1441 aaggtgaagg agaaggccga ccatatgagg gaacacaaac cgcaaaacta aaggtaagtt
     1501 taaacatata tatactaact aaccctgatt atttaaattt tcaggtaact aaaggcggac
     1561 cattaccatt cgcctgggac atcctctctc cacagttcat gtatggaagt aaagcttatg
     1621 ttaaacatcc ggcagatata ccagattatt tgaaactttc attcccggag ggttttaagt
     1681 gggaacgcgt aatgaatttt gaagacggag gagttgttac agtgacgcaa gactcaaggt
     1741 aagtttaaac agttcggtac taactaacca tacatattta aattttcagc ctccaagatg
     1801 gagaatttat ttataaagtc aaacttcgag gaacgaattt cccctcggat ggacctgtta
     1861 tgcagaagaa gactatggga tgggaagctt caagtgaaag aatgtaccct gaagacggtg
     1921 ctcttaaggg agagattaaa caacgtctta aattgaaaga tggaggacat tacgatgctg
     1981 aggtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt caggtgaaga
     2041 caacttacaa agccaaaaaa ccagttcagc tgccaggagc gtacaatgtt aatattaaac
     2101 tggatatcac ctcccacaac gaggattaca ctatcgttga gcaatatgaa agagctgaag
     2161 ggcggcactc gacaggtggc atggatgaat tgtataagac tagtggatcc taaggtaccg
     2221 cggccgcgct gaggggctgg ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2281 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2401 aaccccttgc ggccgcccgg gccgtcgacc aattctcatg tttgacagct tatcatcgaa
     2461 tttctgccat tcatccgctt attatcactt attcaggcgt agcaaccagg cgtttaaggg
     2521 caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt
     2581 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     2641 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     2701 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     2761 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     2821 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     2881 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     2941 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     3001 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     3061 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     3121 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     3181 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     3241 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     3301 cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca
     3361 ccaggattta tttattctgc gaagtgatct tccgtcacag gtatttattc gcgataagct
     3421 catggagcgg cgtaaccgtc gcacaggaag gacagagaaa gcgcggatct gggaagtgac
     3481 ggacagaacg gtcaggacct ggattgggga ggcggttgcc gccgctgctg ctgacggtgt
     3541 gacgttctct gttccggtca caccacatac gttccgccat tcctatgcga tgcacatgct
     3601 gtatgccggt ataccgctga aagttctgca aagcctgatg ggacataagt ccatcagttc
     3661 aacggaagtc tacacgaagg tttttgcgct ggatgtggct gcccggcacc gggtgcagtt
     3721 tgcgatgccg gagtctgatg cggttgcgat gctgaaacaa ttatcctgag aataaatgcc
     3781 ttggccttta tatggaaatg tggaactgag tggatatgct gtttttgtct gttaaacaga
     3841 gaagctggct gttatccact gagaagcgaa cgaaacagtc gggaaaatct cccattatcg
     3901 tagagatccg cattattaat ctcaggagcc tgtgtagcgt ttataggaag tagtgttctg
     3961 tcatgatgcc tgcaagcggt aacgaaaacg atttgaatat gccttcagga acaatagaaa
     4021 tcttcgtgcg gtgttacgtt gaagtggagc ggattatgtc agcaatggac agaacaacct
     4081 aatgaacaca gaaccatgat gtggtctgtc cttttacagc cagtagtgct cgccgcagtc
     4141 gagcgacagg gcgaagccct cggctggttg ccctcgccgc tgggctggcg gccgtctatg
     4201 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccggccgc
     4261 cggcgttgtg gatacctcgc ggaaaacttg gccctcactg acagatgagg ggcggacgtt
     4321 gacacttgag gggccgactc acccggcgcg gcgttgacag atgaggggca ggctcgattt
     4381 cggccggcga cgtggagctg gccagcctcg caaatcggcg aaaacgcctg attttacgcg
     4441 agtttcccac agatgatgtg gacaagcctg gggataagtg ccctgcggta ttgacacttg
     4501 aggggcgcga ctactgacag atgaggggcg cgatccttga cacttgaggg gcagagtgct
     4561 gacagatgag gggcgcacct attgacattt gaggggctgt ccacaggcag aaaatccagc
     4621 atttgcaagg gtttccgccc gtttttcggc caccgctaac ctgtctttta acctgctttt
     4681 aaaccaatat ttataaacct tgtttttaac cagggctgcg ccctgtgcgc gtgaccgcgc
     4741 acgccgaagg ggggtgcccc cccttctcga accctcccgg tcgagtgagc gaggaagcac
     4801 cagggaacag cacttatata ttctgcttac acacgatgcc tgaaaaaact tcccttgggg
     4861 ttatccactt atccacgggg atatttttat aattattttt tttatagttt ttagatcttc
     4921 ttttttagag cgccttgtag gcctttatcc atgctggttc tagagaaggt gttgtgacaa
     4981 attgcccttt cagtgtgaca aatcaccctc aaatgacagt cctgtctgtg acaaattgcc
     5041 cttaaccctg tgacaaattg ccctcagaag aagctgtttt ttcacaaagt tatccctgct
     5101 tattgactct tttttattta gtgtgacaat ctaaaaactt gtcacacttc acatggatct
     5161 gtcatggcgg aaacagcggt tatcaatcac aagaaacgta aaaatagccc gcgaatcgtc
     5221 cagtcaaacg acctcactga ggcggcatat agtctctccc gggatcaaaa acgtatgctg
     5281 tatctgttcg ttgaccagat cagaaaatct gatggcaccc tacaggaaca tgacggtatc
     5341 tgcgagatcc atgttgctaa atatgctgaa atattcggat tgacctctgc ggaagccagt
     5401 aaggatatac ggcaggcatt gaagagtttc gcggggaagg aagtggtttt ttatcgccct
     5461 gaagaggatg ccggcgatga aaaaggctat gaatcttttc cttggtttat caaacgtgcg
     5521 cacagtccat ccagagggct ttacagtgta catatcaacc catatctcat tcccttcttt
     5581 atcgggttac agaaccggtt tacgcagttt cggcttagtg aaacaaaaga aatcaccaat
     5641 ccgtatgcca tgcgtttata cgaatccctg tgtcagtatc gtaagccgga tggctcaggc
     5701 atcgtctctc tgaaaatcga ctggatcata gagcgttacc agctgcctca aagttaccag
     5761 cgtatgcctg acttccgccg ccgcttcctg caggtctgtg ttaatgagat caacagcaga
     5821 actccaatgc gcctctcata cattgagaaa aagaaaggcc gccagacgac tcatatcgta
     5881 ttttccttcc gcgatatcac ttccatgacg acaggatagt ctgagggtta tctgtcacag
     5941 atttgagggt ggttcgtcac atttgttctg acctactgag ggtaatttgt cacagttttg
     6001 ctgtttcctt cagcctgcat ggattttctc atactttttg aactgtaatt tttaaggaag
     6061 ccaaatttga gggcagtttg tcacagttga tttccttctc tttcccttcg tcatgtgacc
     6121 tgatatcggg ggttagttcg tcatcattga tgagggttga ttatcacagt ttattactct
     6181 gaattggcta tccgcgtgtg tacctctacc tggagttttt cccacggtgg atatttcttc
     6241 ttgcgctgag cgtaagagct atctgacaga acagttcttc tttgcttcct cgccagttcg
     6301 ctcgctatgc tcggttacac ggctgcggcg agcgctagtg ataataagtg actgaggtat
     6361 gtgctcttct tatctccttt tgtagtgttg ctcttatttt aaacaacttt gcggtttttt
     6421 gatgactttg cgattttgtt gttgctttgc agtaaattgc aagatttaat aaaaaaacgc
     6481 aaagcaatga ttaaaggatg ttcagaatga aactcatgga aacacttaac cagtgcataa
     6541 acgctggtca tgaaatgacg aaggctatcg ccattgcaca gtttaatgat gacagcccgg
     6601 aagcgaggaa aataacccgg cgctggagaa taggtgaagc agcggattta gttggggttt
     6661 cttctcaggc tatcagagat gccgagaaag cagggcgact accgcacccg gatatggaaa
     6721 ttcgaggacg ggttgagcaa cgtgttggtt atacaattga acaaattaat catatgcgtg
     6781 atgtgtttgg tacgcgattg cgacgtgctg aagacgtatt tccaccggtg atcggggttg
     6841 ctgcccataa aggtggcgtt tacaaaacct cagtttctgt tcatcttgct caggatctgg
     6901 ctctgaaggg gctacgtgtt ttgctcgtgg aaggtaacga cccccaggga acagcctcaa
     6961 tgtatcacgg atgggtacca gatcttcata ttcatgcaga agacactctc ctgcctttct
     7021 atcttgggga aaaggacgat gtcacttatg caataaagcc cacttgctgg ccggggcttg
     7081 acattattcc ttcctgtctg gctctgcacc gtattgaaac tgagttaatg ggcaaatttg
     7141 atgaaggtaa actgcccacc gatccacacc tgatgctccg actggccatt gaaactgttg
     7201 ctcatgacta tgatgtcata gttattgaca gcgcgcctaa cctgggtatc ggcacgatta
     7261 atgtcgtatg tgctgctgat gtgctgattg ttcccacgcc tgctgagttg tttgactaca
     7321 cctccgcact gcagtttttc gatatgcttc gtgatctgct caagaacgtt gatcttaaag
     7381 ggttcgagcc tgatgtacgt attttgctta ccaaatacag caatagtaat ggctctcagt
     7441 ccccgtggat ggaggagcaa attcgggatg cctggggaag catggttcta aaaaatgttg
     7501 tacgtgaaac ggatgaagtt ggtaaaggtc agatccggat gagaactgtt tttgaacagg
     7561 ccattgatca acgctcttca actggtgcct ggagaaatgc tctttctatt tgggaacctg
     7621 tctgcaatga aattttcgat cgtctgatta aaccacgctg ggagattaga taatgaagcg
     7681 tgcgcctgtt attccaaaac atacgctcaa tactcaaccg gttgaagata cttcgttatc
     7741 gacaccagct gccccgatgg tggattcgtt aattgcgcgc gtaggagtaa tggctcgcgg
     7801 taatgccatt actttgcctg tatgtggtcg ggatgtgaag tttactcttg aagtgctccg
     7861 gggtgatagt gttgagaaga cctctcgggt atggtcaggt aatgaacgtg accaggagct
     7921 gcttactgag gacgcactgg atgatctcat cccttctttt ctactgactg gtcaacagac
     7981 accggcgttc ggtcgaagag tatctggtgt catagaaatt gccgatggga gtcgccgtcg
     8041 taaagctgct gcacttaccg aaagtgatta tcgtgttctg gttggcgagc tggatgatga
     8101 gcagatggct gcattatcca gattgggtaa cgattatcgc ccaacaagtg cttatgaacg
     8161 tggtcagcgt tatgcaagcc gattgcagaa tgaatttgct ggaaatattt ctgcgctggc
     8221 tgatgcggaa aatatttcac gtaagattat tacccgctgt atcaacaccg ccaaattgcc
     8281 taaatcagtt gttgctcttt tttctcaccc cggtgaacta tctgcccggt caggtgatgc
     8341 acttcaaaaa gcctttacag ataaagagga attacttaag cagcaggcat ctaaccttca
     8401 tgagcagaaa aaagctgggg tgatatttga agctgaagaa gttatcactc ttttaacttc
     8461 tgtgcttaaa acgtcatctg catcaagaac tagtttaagc tcacgacatc agtttgctcc
     8521 tggagcgaca gtattgtata agggcgataa aatggtgctt aacctggaca ggtctcgtgt
     8581 tccaactgag tgtatagaga aaattgaggc cattcttaag gaacttgaaa agccagcacc
     8641 ctgatgcgac cacgttttag tctacgttta tctgtcttta cttaatgtcc tttgttacag
     8701 gccagaaagc ataactggcc tgaatattct ctctgggccc actgttccac ttgtatcgtc
     8761 ggtctgataa tcagactggg accacggtcc cactcgtatc gtcggtctga ttattagtct
     8821 gggaccacgg tcccactcgt atcgtcggtc tgattattag tctgggacca cggtcccact
     8881 cgtatcgtcg gtctgataat cagactggga ccacggtccc actcgtatcg tcggtctgat
     8941 tattagtctg ggaccatggt cccactcgta tcgtcggtct gattattagt ctgggaccac
     9001 ggtcccactc gtatcgtcgg tctgattatt agtctggaac cacggtccca ctcgtatcgt
     9061 cggtctgatt attagtctgg gaccacggtc ccactcgtat cgtcggtctg attattagtc
     9121 tgggaccacg atcccactcg tgttgtcggt ctgattatcg gtctgggacc acggtcccac
     9181 ttgtattgtc gatcagacta tcagcgtgag actacgattc catcaatgcc tgtcaagggc
     9241 aagtattgac atgtcgtcgt aacctgt
//
