LOCUS       sequence-265902-        6225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of mouse C/EBPbeta in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6225)
  TITLE     Jacob Freidman lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6225)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA-mC/EBPb
FEATURES             Location/Qualifiers
     source          1..6225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..39)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        211..590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        591..794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     745..765
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     839..858
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        839..857
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1292..1301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1792..1809)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1798..2022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2068..2496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2155..2174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2365..2386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2505..2525)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2510..2839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2690..2825
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2752..2771
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2906..3700
                     /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(2960..2979)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3570..3589
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3874..3995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3911..3930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3965..3984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4044..4060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4044..4060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4057..4079)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4068..4084
                     /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(4092..4098,4099..4116,4117..4122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4137..4158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4275..4292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4446..5034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4526..4545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5205..5996,5997..6065))
                     /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     5828..5847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6066..6170)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
       61 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
      121 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
      181 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
      241 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      301 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      361 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      421 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      481 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      541 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      601 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      661 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      721 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      781 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
      841 atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc ttggtacccc
      901 accatggaag tggccaactt ctactacgag cccgactgcc tggcctacgg ggccaaggcg
      961 gcccgcgccg cgccgcgcgc ccccgccgcc gagccggcca ttggcgagca cgagcgcgcc
     1021 atcgacttca gcccctacct ggagccgctc gcgcccgccg cggacttcgc cgcgcccgcg
     1081 cccgcgcacc acgacttcct ctccgacctc ttcgccgacg actacggcgc caagccgagc
     1141 aagaagccgg ccgactacgg ttacgtgagc ctcggccgcg cgggcgccaa ggccgcgccg
     1201 cccgcctgct tcccgccgcc gcctcccgcc gcgctcaagg cggagccggg cttcgaaccc
     1261 gcggactgca agcgcgcgga cgacgcgccc gccatggcgg ccggtttccc gttcgccctg
     1321 cgcgcctacc tgggctacca ggcgacgccg agcggcagca gcggcagcct gtccacgtcg
     1381 tcgtcgtcca gcccgcccgg cacgccgagc cccgccgacg ccaaggccgc gcccgccgcc
     1441 tgcttcgcgg ggccgccggc cgcgcccgcc aaggccaagg ccaagaagac ggtggacaag
     1501 ctgagcgacg agtacaagat gcggcgcgag cgcaacaaca tcgcggtgcg caagagccgc
     1561 gacaaggcca agatgcgcaa cctggagacg cagcacaagg tgctggagct gacggcggag
     1621 aacgagcggc tgcagaagaa ggtggagcag ctgtcgcgag agctcagcac cctgcggaac
     1681 ttgttcaagc agctgcccga gccgctgctg gcctcggcgg gccactgcta ggatatccag
     1741 cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     1801 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1861 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1921 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1981 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2041 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2101 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2161 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2221 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2281 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2341 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2401 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2461 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     2521 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2581 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2641 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2701 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2761 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     2821 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     2881 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     2941 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3001 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3061 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3121 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3181 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3241 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3301 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3361 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3421 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3481 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3541 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3601 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3661 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     3721 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     3781 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     3841 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     3901 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3961 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4021 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4081 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4141 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4201 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4261 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4321 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4381 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4441 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4501 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4561 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4621 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4681 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4741 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4801 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4861 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4921 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4981 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5041 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5101 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5161 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5221 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5281 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5341 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5401 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5461 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5521 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5581 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5641 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5701 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5761 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5821 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5881 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5941 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6001 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6061 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6121 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6181 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatc
//
