LOCUS       sequence-6883-ex        7742 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 7742)
  AUTHORS   Lepourcelet M, Shivdasani RA
  TITLE     Characterization of a novel mammalian Groucho isoform and its role 
            in transcriptional regulation.
  JOURNAL   J Biol Chem. 2002 Dec 6. 277(49):47732-40.
  PUBMED    12359720
REFERENCE   2  (bases 1 to 7742)
  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: "J Biol 
            Chem. 2002 Dec 6. 277(49):47732-40."
COMMENT     Sequence Label: p178 Grg-1L Myc His
FEATURES             Location/Qualifiers
     source          1..7742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3245..3274
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3290..3307
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3330..3347)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3336..3560
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3606..4034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3693..3712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3903..3924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4043..4063)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4048..4378
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4229..4364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4291..4310
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4445..5239
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4499..4518)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5109..5128
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5415..5536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5452..5471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5506..5525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5585..5601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5585..5601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5598..5620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5609..5625
                     /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(5633..5639,5640..5657,5658..5663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5678..5699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5816..5833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5987..6575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6067..6086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6746..7537,7538..7606))
                     /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     7369..7388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7607..7711)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccatgtt cccgcagagc cggcacccaa cgccgcacca
      961 agctgcaggc cagcccttta agttcactat cccggagtct ctggaccgga ttaaagagga
     1021 attccagttc ctgcaggcgc agtatcacag tcttaaattg gagtgtggga aactggcaag
     1081 tgaaaagaca gaaatgcaga ggcactacgt gatgtattat gaaatgtcat atggattaaa
     1141 cattgaaatg cacaaacaga ctgaaatcgc caagagattg aacaccattt gtgcccaagt
     1201 catcccattt ctgtctcagg aacatcaaca acaggtggcc caggctgtgg aacgtgccaa
     1261 acaggtgacc atggcagagt tgaatgccat catcgggcag cagcagttgc aagctcagca
     1321 tctctcccat ggccatggac ccccagtacc tctcacgcct cacccttcag gacttcagcc
     1381 tcctggcatc ccgccccttg ggggcagtgc cagccttctc gcgctgtcta gtgctctgag
     1441 tgggcagtct cacttggcaa taaaagatga caagaagcac catgatgcag agcgccacag
     1501 agacagagag cctggcacga gtaattccct cttggtccca gacagcctaa gaagcacaga
     1561 taagcgcaga aatggtccag agttttccag tgacatcaaa aaaaggaagg tggatgataa
     1621 ggataactat gacagtgatg gggacaagag tgatgacaac ttaattgtgg atgtgtctaa
     1681 cgaggaccct tcttcttcac atgcaagccc cacacactca ccccgggaaa acggaattga
     1741 caaaaaccgt ctgctgaaga aagatgcttc aggtagccca gcatccacag cctcctctgg
     1801 aagttcctct tccctgaaat ccaaagaagt gagcctgcat gaaaaagcca acactcctgt
     1861 tctgaaatcc agcacaccga cgcctcggag cgacatgcca accccaggca ccagcgctac
     1921 tccaggcctc cgtccaggtc tcggcaaacc tccagccatg gaaccccttg tcaaccaagc
     1981 agcagctggc ctgaggacgc ccctggcggt gcctggccca taccctgccc cctttggcat
     2041 ggtgccccat gcgggcatga acggagagct gaccagccct ggtgctgcct atgcaggtct
     2101 acacagcatg tctccacaga tgagcgctgc agctgctgca gctgctgctg ctgtggtggc
     2161 ctatgggcgc tccccaatgg ttggttttga tcctcctcct cacatgagag taccttctat
     2221 cccccccaac ctggcaggaa tacctggagg gaaaccagca tactccttcc atgttactgc
     2281 tgatggccaa atgcagcctg ttccttttcc ccccgatgcc ctcattggac ctgggattcc
     2341 ccgacatgct cggcagatca acaccctcaa ccacggggag gtggtgtgcg cagtgaccat
     2401 cagcaacccc acaaggcacg tgtacacagg tggcaagggc tgcgttaagg tgtgggacat
     2461 cagccacccc ggcaacaaga gccccgtctc tcagctggat tgtctgaata gagataacta
     2521 catccgttcc tgtaaattgc tacctgatgg ctgtactctc atagtgggag gggaagccag
     2581 tactctatcc atctgggact tggcagctcc aactccccgc ataaaggcag agctgacatc
     2641 ctcagcccct gcctgttatg ctctggccat cagccccgac tccaaggtct gcttctcatg
     2701 ctgcagtgac ggcaacatcg cagtgtggga cctgcacaac cagaccctgg tgaggcaatt
     2761 ccagggccac acagacggag ccagctgtat tgacatttct aatgatggca ccaagctctg
     2821 gacaggcggt ttagacaaca ctgtgaggtc ctgggacctg agagaaggga ggcagctgca
     2881 gcagcatgac ttcacttcac agatcttctc tttgggatat tgcccaactg gggagtggct
     2941 tgctgtgggc atggaaagca gcaacgtgga agttctgcat gtgaacaagc ctgacaagta
     3001 ccagctgcac ctccacgaga gctgcgtgct gtccctaaag tttgcttatt gtggcaaatg
     3061 gtttgtgagt actggaaaag acaacctcct aaatgcctgg cggacaccat atggagccag
     3121 catcttccag tccaaagaat cttcatcagt gcttagctgt gacatctctg tggatgataa
     3181 atacatagtc actggctcag gggacaaaaa ggctaccgtt tatgaggtca tctacccgcg
     3241 gttcgaacaa aaactcatct cagaagagga tctgaatatg cataccggtc atcatcacca
     3301 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3361 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3421 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3481 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3541 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3601 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3661 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3721 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     3781 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3841 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3901 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3961 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4021 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4081 tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4141 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4201 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4261 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4321 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4381 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     4441 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4501 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4561 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4621 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4681 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4741 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4801 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4861 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4921 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4981 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5041 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5101 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5161 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5221 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     5281 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5341 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5401 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5521 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5581 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5641 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5701 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5761 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5821 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5881 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5941 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6001 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6061 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6121 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6181 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6241 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6301 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6361 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6421 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6481 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6541 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6601 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6661 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6721 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6781 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6841 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6901 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6961 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7021 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7081 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7141 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7201 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7261 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7321 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7381 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7441 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7501 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7561 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7621 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7681 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7741 tc
//
