LOCUS       sequence-223078-        8134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recombination substrate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8134)
  AUTHORS   Lieber MR, Hesse JE, Lewis S, Bosma GC, Rosenberg N, Mizuuchi K, 
            Bosma MJ, Gellert M
  TITLE     The defect in murine severe combined immune deficiency: joining of 
            signal sequences but not coding segments in V(D)J recombination.
  JOURNAL   Cell. 1988 Oct 7. 55(1):7-16.
  PUBMED    3167977
REFERENCE   2  (bases 1 to 8134)
  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: "Cell. 1988 
            Oct 7. 55(1):7-16."
COMMENT     Sequence Label: pJH298
FEATURES             Location/Qualifiers
     source          1..8134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      836..1349
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(923..942)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1133..1154
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1602..2261)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2176..2195
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(2430..2464)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     complement(2675..2691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2675..2691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2688..2710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2699..2715
                     /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)."
     promoter        complement(join(2723..2729,2730..2747,2748..2753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2768..2789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2906..2923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3077..3665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3157..3176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3836..4627,4628..4696))
                     /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     4459..4478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4697..4801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4869..4887
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4925..4947)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5047..5066
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5260..5282
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5274..5291
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5275..5291
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(7077..7094)
                     /codon_start=1
                     /product="epitope tag from middle T antigen of mouse 
                     polyoma virus"
                     /label=Glu-Glu tag
                     /translation="EYMPME"
ORIGIN
        1 ggggcttaat tatatcaatt tgggtggccc cacagcgcct ccaaggcgcc agtcctgttt
       61 tgacaagttg cctctggaag cctctacaat gcctctcttc tttttctcca gagtaaagcg
      121 gaggccaggg gcccccggcc tctgcttaat actaaaaaaa acagctagtg acttcctgct
      181 taactgactc actgccctct ggagggcagt tgcctagcaa ctaattaaaa gaggatgtcg
      241 cacggccagc tgcggtcagc tagtgacttc ctgcttaact gactcactgc cctctggagg
      301 gcagttgcct agcaactaat taaaagagga tgtgactcac ggccagcagc tgcggtcagc
      361 tagtgactca cggccagctg cggtcagcta gtgacttcct gcttaactga cttgacattt
      421 tctattttaa gagtcgggag gaaaattact gtgttggagg ccctccgcca tcttctgaag
      481 ctgatcaagt aagtgaattt tcaaaatggg agccgcactg actattctag tagacctcat
      541 cgagggatta gctgaagtgt ctacccttac gggactctcg gcagaagcta ttttatctgg
      601 agaagccctc gctgcccttg atggcgaaat tacagctctg actttggagg gtgtaatgag
      661 ttcggagaca gccctagcaa ctatgggtat ttcagaggag gtgtatgggt ttgtcagtac
      721 tgtgcctgtg tttgtaaatc gaacagcggg ggctatatgg ctgatgcaga cagttcaagg
      781 tgcctctact atttccctag cagatctgag cttgcatgcc tgcaggtcga tccccacgcg
      841 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
      901 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
      961 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1021 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1081 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1141 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1201 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1261 aattttaaca aaatattaac gcttacaatt taaatatttg cttatacaat cttcctgttt
     1321 ttggggcttt tctgattatc aaccggggtg ggtaccgagc tcgaattcag atctgctagg
     1381 aatacagcgg tacctacaca acgaagaggt ccctactgta aatagaaata tggcgttgat
     1441 accatggcgg gatcaattcg agctcgcccg gggacggatc gactctaggg gtaccgagct
     1501 cgaattccct ccggcgaatt tctgccattc atccgcttat tatcacttat tcaggcgtag
     1561 caaccaggcg tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac
     1621 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     1681 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     1741 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     1801 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     1861 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     1921 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     1981 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     2041 cggaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     2101 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     2161 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     2221 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     2281 gaaaatctcg ccaagctgat ccccggggat cccgagggtt tttgtacagc cagacagtgg
     2341 agtactacca ctgtgggatc ctctcatcga tgagaggatc gacgacgaca tggctcgatt
     2401 ggcgcgacaa gttgctgcga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     2461 tgaacaaatc cagatggagt tctgaggtca ttactggaga cctgccgagt ggttcagcag
     2521 gtggaagagg gactggattc caaagttctc aatgctgctt gctgttcttg aatggggggt
     2581 cgttgatccc ccatcgatga gagtcgacca cagtgctaca gactggaaca aaaaccctgc
     2641 agtcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2701 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2761 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2821 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2881 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4801 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4861 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     4921 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4981 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     5041 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     5101 acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact
     5161 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     5221 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     5281 cgacggccag tgaattgacc gctttggaaa aacaaagact gtatttcctg gaaattaatg
     5341 tttattcaat aaactgtgta ttcagctata ttcacatatg tccttcaagg ggtcgtcgcc
     5401 ttcttgcaca ttacaaagta tatcacaaaa caaactgtag gagcactcac tagcaaactg
     5461 gtcacgaacc tccttcacaa gaccctgaat atcaggatca aatacatctg aagtgaaatt
     5521 ccatatgagt aatagggcaa gggtatctcc actctgaata attctttccc tttgcaaaaa
     5581 ttcacacttt tcaagagatt gggccagatg gggtttgcag gtgaaatcca acaccatgtg
     5641 aaaccgggcc catactgttt gtggtaggag atattcattc attgtacaca cacagggtgg
     5701 aaagagttgt gaccttttgt tgctgtgctt tttttctaga ttgaccttta cactcccatc
     5761 caggtagtcc ctgagattat ctagattagc cactcccatc cctggctgca gttgtttgtt
     5821 caaggctatt tgacccttta catcttcaaa acagaccaca aactggtcct gtgccacacc
     5881 aagctcaaaa gcaagtttat ctgcagggca atttatgttg agagacttgc ctcctaacag
     5941 gctaataagc gcggctgcta ggcctgtctt tcctgaatta actggtcctc taaaaagtat
     6001 gttgcgtcgt tttggcacat tttctgttag cagcttaagc atcttaaaca gggtttgagg
     6061 aaagtcctct aaaagacact ggtaccaagc gacacctgct aggtagagta gagcatcgga
     6121 ggagccaagt tcttttagcc tgaggagaga ctgttgcaat ctctccttta atagctggct
     6181 gcgggtacac tctactaatt tcagtctcct ggatgccaag actccgtcag ctgcctgctg
     6241 acagattgtc ttttgagctt tacaattcag gaagaggtct gcattctctg catgctttct
     6301 atggtttttc caatgtattt ggaggcgggt ttcctctttg ctacacttta tgcatgaagg
     6361 aacctctttg gcaaaatcaa gataataccc cataattaac aggggatcat caaggttgtt
     6421 ttctagtgca aagtctgcta ccataatcca gtctactgct ttctgttctt ctggctcgtc
     6481 tgtaaactcg aattggtgga ggcctggctt attttctgtt attaactgaa atggtgctgc
     6541 ggttacaact tgatagcatt ccataggctt ggtgactgcc ttacacatta ggaagctgac
     6601 gctgcaaagc ttagagcaat aattcttaac tgctgaaacc ctgtgcttag tcatagttag
     6661 aaagaacagc atgcccccct cctcataatg gaccaggcat ttgaattcgg gcctgaactt
     6721 ccctatggta tcatataatt gtttgcattt ctctttggtg gagtatacta gaaatgccgg
     6781 gaacgtttta ttagaataaa tagcatgaga caaataccca gtaaggctgc taggaaagtc
     6841 actaggagca gggtcctccc tagccttctt aggtggcgtt gcattgaatg agctctgaga
     6901 gcagctctgt gagtagctct cattctctga ctcgcatcta ggatcggatc gaaatgagcc
     6961 ctctgcaaat cccgaagaat cagaccctcc catggactca gacccgccct gggaatgata
     7021 ggggggggga tgagctgggg tacttgttcc tccggtagga tgtccaaata cagatcctcc
     7081 attggcatgt actcctcctc ctcctcctcc agtggctcca gctggtcttg gtcgctttct
     7141 ggatacagga gttctggggg ggtccgggga tatgctgtca tcgggctcag caacacaagg
     7201 atttcgtctt cttccagcag tatgtgcggg tgtatctgtt ggagaactcg ggttggggga
     7261 taggagtggc tcctcataac agaataggtc gggttgctca gaagactcgg cagtcttagg
     7321 cggcgactgg tagctggggt cccactttct gaagaaatca ttccattccc ttgtgaagta
     7381 cttgcttcca tagctgaatg accagtagtc atcgtgtagt ggactgtggc cgctctcctc
     7441 agttcctcgc tccgccgttc tagggggaga gccctgtaag aatataggcc gtaaataccc
     7501 acccgccctc ttgatactta cttggattat acacgctgtg cacatccagg tccagccagt
     7561 ctataggcat gcttgcaatg aagtctgcat acaggttcag cacatctcgg gttggtgttc
     7621 caaaccattg catgtaacat tcaagacaaa aacattctcc aagtactagg cacctggcat
     7681 cacatttgtc tttgagctct ctatgttgct ttctaagcag gcataatata caactacatg
     7741 agctgtattt aacatttact aggcaggtaa gaggcattct gcagaaccgc tggtagtatc
     7801 tatcaccaaa ggtgtctttg gtacttagat tccacccatc cgcatgtagc cttcttacct
     7861 ggaagccggt tcctcctaga ttcattctca gattgtatac ttcagtttta aatgttcccc
     7921 agagactgtt caattcctgc attaaggcat ggcttccacc tttgtctggg tgcagtagca
     7981 gtgactgctg cttatatgcc tgctgcattc ttccaaaatc cccccatagt tgtctgggaa
     8041 gttttagaag ttctagcagc ctttctttgt cagctctgct cagaactcta tccatgatgg
     8101 tggtgaggct gaaatgaggc gggaagaggc ggtt
//
