LOCUS       sequence-177731-        7968 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 7968)
  AUTHORS   Johnson TK, Schweppe RE, Septer J, Lewis RE
  TITLE     Phosphorylation of B-Myb regulates its transactivation potential and
            DNA binding.
  JOURNAL   J Biol Chem. 1999 Dec 17. 274(51):36741-9.
  PUBMED    10593981
REFERENCE   2  (bases 1 to 7968)
  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. 1999 Dec 17. 274(51):36741-9."
COMMENT     Sequence Label: pCDNA3 WT B-Myb
FEATURES             Location/Qualifiers
     source          1..7968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        233..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        613..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     767..787
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     861..880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        861..879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3519..3537)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3520..3537)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3557..3574)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3563..3787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3833..4261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3920..3939)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4130..4151
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4270..4290)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4275..4604
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4455..4590
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4517..4536
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4671..5465
                     /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(4725..4744)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5335..5354
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5639..5760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5676..5695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5730..5749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5809..5825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5809..5825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5822..5844)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5833..5849
                     /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(5857..5863,5864..5881,5882..5887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5902..5923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6040..6057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6211..6799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6291..6310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6970..7761,7762..7830))
                     /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     7593..7612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7831..7935)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
       61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga
      901 gctcggatcc atgtctcggc ggacgcgctg cgaggatctg gatgagctgc actaccagga
      961 cacagattca gatgtgccgg agcagaggga tagcaagtgc aaggtcaaat ggacccatga
     1021 ggaggacgag cagctgaggg ccctggtgag gcagtttgga cagcaggact ggaagttcct
     1081 ggccagccac ttccctaacc gcactgacca gcaatgccag tacaggtggc tgagagtttt
     1141 gaatccagac cttgtcaagg ggccatggac caaagaggaa gaccaaaaag tcatcgagct
     1201 ggttaagaag tatggcacaa agcagtggac actgattgcc aagcacctga agggccggct
     1261 ggggaagcag tgccgtgaac gctggcacaa ccacctcaac cctgaggtga agaagtcttg
     1321 ctggaccgag gaggaggacc gcatcatctg cgaggcccac aaggtgctgg gcaaccgctg
     1381 ggccgagatc gccaagatgt tgccagggag gacagacaat gctgtgaaga atcactggaa
     1441 ctctaccatc aaaaggaagg tggacacagg aggcttcttg agcgagtcca aagactgcaa
     1501 gcccccagtg tacttgctgc tggagctcga ggacaaggac ggcctccaga gtgcccagcc
     1561 cacggaaggc cagggaagtc ttctgaccaa ctggccctcc gtccctccta ccataaagga
     1621 ggaggaaaac agtgaggagg aacttgcagc agccaccaca tcgaaggaac aggagcccat
     1681 cggtacagat ctggacgcag tgcgaacacc agagcccttg gaggaattcc cgaagcgtga
     1741 ggaccaggaa ggctccccac cagaaacgag cctgccttac aagtgggtgg tggaggcagc
     1801 taacctcctc atccccgctg tgggttctag cctctctgaa gccctggact tgatcgagtc
     1861 ggaccctgat gcttggtgtg acctgagtaa atttgacctc cctgaggaac catctgcaga
     1921 ggacagtatc aacaacagcc tagtgcagct gcaagcgtca catcagcagc aagtcctgcc
     1981 accccgccag ccttccgccc tggtgcccag tgtgaccgag taccgcctgg atggccacac
     2041 catctcagac ctgagccgga gcagccgggg cgagctgatc cccatctccc ccagcactga
     2101 agtcgggggc tctggcattg gcacaccgcc ctctgtgctc aagcggcaga ggaagaggcg
     2161 tgtggctctg tcccctgtca ctgagaatag caccagtctg tccttcctgg attcctgtaa
     2221 cagcctcacg cccaagagca cacctgttaa gaccctgccc ttctcgccct cccagtttct
     2281 gaacttctgg aacaaacagg acacattgga gctggagagc ccctcgctga catccacccc
     2341 agtgtgcagc cagaaggtgg tggtcaccac accactgcac cgggacaaga cacccctgca
     2401 ccagaaacat gctgcgtttg taaccccaga tcagaagtac tccatggaca acactcccca
     2461 cacgccaacc ccgttcaaga acgccctgga gaagtacgga cccctgaagc ccctgccaca
     2521 gaccccgcac ctggaggagg acttgaagga ggtgctgcgt tctgaggctg gcatcgaact
     2581 catcatcgag gacgacatca ggcccgagaa gcagaagagg aagcctgggc tgcggcggag
     2641 ccccatcaag aaagtccgga agtctctggc tcttgacatt gtggatgagg atgtgaagct
     2701 gatgatgtcc acactgccca agtctctatc cttgccgaca actgcccctt caaactcttc
     2761 cagcctcacc ctgtcaggta tcaaagaaga caacagcttg ctcaaccagg gcttcttgca
     2821 ggccaagccc gagaaggcag cagtggccca gaagccccga agccacttca cgacacctgc
     2881 ccctatgtcc agtgcctgga agacggtggc ctgcgggggg accagggacc agcttttcat
     2941 gcaggagaaa gcccggcagc tcctgggccg cctgaagccc agccacacat ctcggaccct
     3001 catcttgtcc tgaggtgttg agggtgtcac gagcccattc tcatgtttac aggggttgtg
     3061 ggggcagagg gggtctgtga atctgagagt cattcaggtg acctcctgca gggagccttc
     3121 tgccaccagc ccctccccag actctcaggt ggaggcaaca gggccatgtg ctgccctgtt
     3181 gccgagccca gctgtgggcg gctcctggtg ctaacaacaa agttccactt ccaggtctgc
     3241 ctggttccct ccccaaggcc acagggagct ccgtcagctt ctcccaagcc cacgtcaggc
     3301 ctggcctcat ctcagaccct gcttaggatg ggggatgtgg ccaggggtgc tcctgtgctc
     3361 accctctctt ggtgcatttt tttggaagaa taaaattgcc tgcggccgcg aattcctgca
     3421 gcccggggga tccactagta acggccgcca gtgtgctgga attctgcaga tatccatcac
     3481 actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct
     3541 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3601 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3661 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3721 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3781 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3841 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3901 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3961 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4021 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4081 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4141 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4201 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4261 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4321 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4381 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4441 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4501 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4561 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4621 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4681 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4741 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4801 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4861 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4921 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4981 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5041 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5101 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5161 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5221 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5281 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5341 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5401 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5461 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5521 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5581 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5641 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5701 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5761 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5821 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5881 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5941 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6001 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6061 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6121 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6181 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6241 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6301 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6361 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6421 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6481 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6541 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6601 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6661 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6721 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6781 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6841 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6901 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6961 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7021 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7081 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7141 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7201 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7261 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7321 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7381 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7441 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7501 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7561 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7621 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7681 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7741 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7801 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7861 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7921 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcga
//
