LOCUS       sequence-56934-e        9261 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 9261)
  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 9261)
  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: pNH040
FEATURES             Location/Qualifiers
     source          1..9261
                     /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
     primer_bind     complement(1368..1396)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2144..2164
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(join(2252..2258,2259..2276,2277..2282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2297..2318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(2538..3197)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3112..3131
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3198..3300)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4158..4772
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      4249..4780
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      4848..5067
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             5158..5913
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    5916..6166
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             6492..7667
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             7667..8638
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    8711..9184
                     /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 agtaaaggag aagaactttt cactggagtt gtcccaattc
     1381 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
     1441 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg
     1501 ttccatgggt aagtttaaac atatatatac taactaaccc tgattattta aattttcagc
     1561 caacacttgt cactactttc tgttatggtg ttcaatgctt ctcgagatac ccagatcata
     1621 tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     1681 tatttttcaa agatgacggg aactacaaga cacgtaagtt taaacagttc ggtactaact
     1741 aaccatacat atttaaattt tcaggtgctg aagtcaagtt tgaaggtgat acccttgtta
     1801 atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat
     1861 tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa
     1921 tcaaagttgt aagtttaaac atgattttac taactaacta atctgattta aattttcaga
     1981 acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac
     2041 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac
     2101 aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg
     2161 taacagctgc tgggattaca catggcatgg atgaactata caaatagggt accgcggccg
     2221 cgctgagggg ctggccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2281 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2341 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgaacccc
     2401 ttgcggccgc ccgggccgtc gaccaattct catgtttgac agcttatcat cgaatttctg
     2461 ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa
     2521 taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     2581 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     2641 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     2701 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     2761 gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     2821 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     2881 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     2941 tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc
     3001 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     3061 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     3121 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     3181 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     3241 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca
     3301 acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga
     3361 tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcgcgata agctcatgga
     3421 gcggcgtaac cgtcgcacag gaaggacaga gaaagcgcgg atctgggaag tgacggacag
     3481 aacggtcagg acctggattg gggaggcggt tgccgccgct gctgctgacg gtgtgacgtt
     3541 ctctgttccg gtcacaccac atacgttccg ccattcctat gcgatgcaca tgctgtatgc
     3601 cggtataccg ctgaaagttc tgcaaagcct gatgggacat aagtccatca gttcaacgga
     3661 agtctacacg aaggtttttg cgctggatgt ggctgcccgg caccgggtgc agtttgcgat
     3721 gccggagtct gatgcggttg cgatgctgaa acaattatcc tgagaataaa tgccttggcc
     3781 tttatatgga aatgtggaac tgagtggata tgctgttttt gtctgttaaa cagagaagct
     3841 ggctgttatc cactgagaag cgaacgaaac agtcgggaaa atctcccatt atcgtagaga
     3901 tccgcattat taatctcagg agcctgtgta gcgtttatag gaagtagtgt tctgtcatga
     3961 tgcctgcaag cggtaacgaa aacgatttga atatgccttc aggaacaata gaaatcttcg
     4021 tgcggtgtta cgttgaagtg gagcggatta tgtcagcaat ggacagaaca acctaatgaa
     4081 cacagaacca tgatgtggtc tgtcctttta cagccagtag tgctcgccgc agtcgagcga
     4141 cagggcgaag ccctcggctg gttgccctcg ccgctgggct ggcggccgtc tatggccctg
     4201 caaacgcgcc agaaacgccg tcgaagccgt gtgcgagaca ccgcggccgg ccgccggcgt
     4261 tgtggatacc tcgcggaaaa cttggccctc actgacagat gaggggcgga cgttgacact
     4321 tgaggggccg actcacccgg cgcggcgttg acagatgagg ggcaggctcg atttcggccg
     4381 gcgacgtgga gctggccagc ctcgcaaatc ggcgaaaacg cctgatttta cgcgagtttc
     4441 ccacagatga tgtggacaag cctggggata agtgccctgc ggtattgaca cttgaggggc
     4501 gcgactactg acagatgagg ggcgcgatcc ttgacacttg aggggcagag tgctgacaga
     4561 tgaggggcgc acctattgac atttgagggg ctgtccacag gcagaaaatc cagcatttgc
     4621 aagggtttcc gcccgttttt cggccaccgc taacctgtct tttaacctgc ttttaaacca
     4681 atatttataa accttgtttt taaccagggc tgcgccctgt gcgcgtgacc gcgcacgccg
     4741 aaggggggtg cccccccttc tcgaaccctc ccggtcgagt gagcgaggaa gcaccaggga
     4801 acagcactta tatattctgc ttacacacga tgcctgaaaa aacttccctt ggggttatcc
     4861 acttatccac ggggatattt ttataattat tttttttata gtttttagat cttctttttt
     4921 agagcgcctt gtaggccttt atccatgctg gttctagaga aggtgttgtg acaaattgcc
     4981 ctttcagtgt gacaaatcac cctcaaatga cagtcctgtc tgtgacaaat tgcccttaac
     5041 cctgtgacaa attgccctca gaagaagctg ttttttcaca aagttatccc tgcttattga
     5101 ctctttttta tttagtgtga caatctaaaa acttgtcaca cttcacatgg atctgtcatg
     5161 gcggaaacag cggttatcaa tcacaagaaa cgtaaaaata gcccgcgaat cgtccagtca
     5221 aacgacctca ctgaggcggc atatagtctc tcccgggatc aaaaacgtat gctgtatctg
     5281 ttcgttgacc agatcagaaa atctgatggc accctacagg aacatgacgg tatctgcgag
     5341 atccatgttg ctaaatatgc tgaaatattc ggattgacct ctgcggaagc cagtaaggat
     5401 atacggcagg cattgaagag tttcgcgggg aaggaagtgg ttttttatcg ccctgaagag
     5461 gatgccggcg atgaaaaagg ctatgaatct tttccttggt ttatcaaacg tgcgcacagt
     5521 ccatccagag ggctttacag tgtacatatc aacccatatc tcattccctt ctttatcggg
     5581 ttacagaacc ggtttacgca gtttcggctt agtgaaacaa aagaaatcac caatccgtat
     5641 gccatgcgtt tatacgaatc cctgtgtcag tatcgtaagc cggatggctc aggcatcgtc
     5701 tctctgaaaa tcgactggat catagagcgt taccagctgc ctcaaagtta ccagcgtatg
     5761 cctgacttcc gccgccgctt cctgcaggtc tgtgttaatg agatcaacag cagaactcca
     5821 atgcgcctct catacattga gaaaaagaaa ggccgccaga cgactcatat cgtattttcc
     5881 ttccgcgata tcacttccat gacgacagga tagtctgagg gttatctgtc acagatttga
     5941 gggtggttcg tcacatttgt tctgacctac tgagggtaat ttgtcacagt tttgctgttt
     6001 ccttcagcct gcatggattt tctcatactt tttgaactgt aatttttaag gaagccaaat
     6061 ttgagggcag tttgtcacag ttgatttcct tctctttccc ttcgtcatgt gacctgatat
     6121 cgggggttag ttcgtcatca ttgatgaggg ttgattatca cagtttatta ctctgaattg
     6181 gctatccgcg tgtgtacctc tacctggagt ttttcccacg gtggatattt cttcttgcgc
     6241 tgagcgtaag agctatctga cagaacagtt cttctttgct tcctcgccag ttcgctcgct
     6301 atgctcggtt acacggctgc ggcgagcgct agtgataata agtgactgag gtatgtgctc
     6361 ttcttatctc cttttgtagt gttgctctta ttttaaacaa ctttgcggtt ttttgatgac
     6421 tttgcgattt tgttgttgct ttgcagtaaa ttgcaagatt taataaaaaa acgcaaagca
     6481 atgattaaag gatgttcaga atgaaactca tggaaacact taaccagtgc ataaacgctg
     6541 gtcatgaaat gacgaaggct atcgccattg cacagtttaa tgatgacagc ccggaagcga
     6601 ggaaaataac ccggcgctgg agaataggtg aagcagcgga tttagttggg gtttcttctc
     6661 aggctatcag agatgccgag aaagcagggc gactaccgca cccggatatg gaaattcgag
     6721 gacgggttga gcaacgtgtt ggttatacaa ttgaacaaat taatcatatg cgtgatgtgt
     6781 ttggtacgcg attgcgacgt gctgaagacg tatttccacc ggtgatcggg gttgctgccc
     6841 ataaaggtgg cgtttacaaa acctcagttt ctgttcatct tgctcaggat ctggctctga
     6901 aggggctacg tgttttgctc gtggaaggta acgaccccca gggaacagcc tcaatgtatc
     6961 acggatgggt accagatctt catattcatg cagaagacac tctcctgcct ttctatcttg
     7021 gggaaaagga cgatgtcact tatgcaataa agcccacttg ctggccgggg cttgacatta
     7081 ttccttcctg tctggctctg caccgtattg aaactgagtt aatgggcaaa tttgatgaag
     7141 gtaaactgcc caccgatcca cacctgatgc tccgactggc cattgaaact gttgctcatg
     7201 actatgatgt catagttatt gacagcgcgc ctaacctggg tatcggcacg attaatgtcg
     7261 tatgtgctgc tgatgtgctg attgttccca cgcctgctga gttgtttgac tacacctccg
     7321 cactgcagtt tttcgatatg cttcgtgatc tgctcaagaa cgttgatctt aaagggttcg
     7381 agcctgatgt acgtattttg cttaccaaat acagcaatag taatggctct cagtccccgt
     7441 ggatggagga gcaaattcgg gatgcctggg gaagcatggt tctaaaaaat gttgtacgtg
     7501 aaacggatga agttggtaaa ggtcagatcc ggatgagaac tgtttttgaa caggccattg
     7561 atcaacgctc ttcaactggt gcctggagaa atgctctttc tatttgggaa cctgtctgca
     7621 atgaaatttt cgatcgtctg attaaaccac gctgggagat tagataatga agcgtgcgcc
     7681 tgttattcca aaacatacgc tcaatactca accggttgaa gatacttcgt tatcgacacc
     7741 agctgccccg atggtggatt cgttaattgc gcgcgtagga gtaatggctc gcggtaatgc
     7801 cattactttg cctgtatgtg gtcgggatgt gaagtttact cttgaagtgc tccggggtga
     7861 tagtgttgag aagacctctc gggtatggtc aggtaatgaa cgtgaccagg agctgcttac
     7921 tgaggacgca ctggatgatc tcatcccttc ttttctactg actggtcaac agacaccggc
     7981 gttcggtcga agagtatctg gtgtcataga aattgccgat gggagtcgcc gtcgtaaagc
     8041 tgctgcactt accgaaagtg attatcgtgt tctggttggc gagctggatg atgagcagat
     8101 ggctgcatta tccagattgg gtaacgatta tcgcccaaca agtgcttatg aacgtggtca
     8161 gcgttatgca agccgattgc agaatgaatt tgctggaaat atttctgcgc tggctgatgc
     8221 ggaaaatatt tcacgtaaga ttattacccg ctgtatcaac accgccaaat tgcctaaatc
     8281 agttgttgct cttttttctc accccggtga actatctgcc cggtcaggtg atgcacttca
     8341 aaaagccttt acagataaag aggaattact taagcagcag gcatctaacc ttcatgagca
     8401 gaaaaaagct ggggtgatat ttgaagctga agaagttatc actcttttaa cttctgtgct
     8461 taaaacgtca tctgcatcaa gaactagttt aagctcacga catcagtttg ctcctggagc
     8521 gacagtattg tataagggcg ataaaatggt gcttaacctg gacaggtctc gtgttccaac
     8581 tgagtgtata gagaaaattg aggccattct taaggaactt gaaaagccag caccctgatg
     8641 cgaccacgtt ttagtctacg tttatctgtc tttacttaat gtcctttgtt acaggccaga
     8701 aagcataact ggcctgaata ttctctctgg gcccactgtt ccacttgtat cgtcggtctg
     8761 ataatcagac tgggaccacg gtcccactcg tatcgtcggt ctgattatta gtctgggacc
     8821 acggtcccac tcgtatcgtc ggtctgatta ttagtctggg accacggtcc cactcgtatc
     8881 gtcggtctga taatcagact gggaccacgg tcccactcgt atcgtcggtc tgattattag
     8941 tctgggacca tggtcccact cgtatcgtcg gtctgattat tagtctggga ccacggtccc
     9001 actcgtatcg tcggtctgat tattagtctg gaaccacggt cccactcgta tcgtcggtct
     9061 gattattagt ctgggaccac ggtcccactc gtatcgtcgg tctgattatt agtctgggac
     9121 cacgatccca ctcgtgttgt cggtctgatt atcggtctgg gaccacggtc ccacttgtat
     9181 tgtcgatcag actatcagcg tgagactacg attccatcaa tgcctgtcaa gggcaagtat
     9241 tgacatgtcg tcgtaacctg t
//
