LOCUS       sequence-148204-        5824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged human Erk1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5824)
  TITLE     cobb lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5824)
  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            "Unpublished"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: pFLAG-CMV-hErk1
FEATURES             Location/Qualifiers
     source          1..5824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     235..254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        235..253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(292..621)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      306..441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     606..626
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    650..1272
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1054..1073
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2441..2506)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(2536..2739)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2565..2585)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2740..3119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3280..3297)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3280..3296)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3289..3311)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3437..3892)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3574..3595)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3786..3805
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3919..4023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4024..4092,4093..4884)
                     /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     complement(4242..4261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5055..5643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5544..5563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5797..5814
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttaatac
      241 gactcactat agggaattaa ctttctggtt tttcagttcc tcgaggagct ttttgcaaaa
      301 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
      361 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
      421 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      481 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      541 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      601 tccacaccct aactgacaca cattccacag ccaagctggc cgcgtaccca attcaacagg
      661 catctactga gtggacccaa cgcatgagag gacagtgcca agcaagcaac tcaaatgtcc
      721 caccggttgg gcatggccag gtagcctatg ctgtgtctgg acgtcctcct gctggtatag
      781 ttattttaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa
      841 tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa
      901 tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc
      961 ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga
     1021 gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca
     1081 ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat
     1141 aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag
     1201 gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca
     1261 gggatgccac ccgggatcct ctctaggggg cctccagcac tccgggctgg aagcgtgctg
     1321 tctcctggaa gatgagctcc ttcagccgct ccttaggtag gtcatccagc tccatggcga
     1381 aggtgaaggg ctcctcggcc actggctcat ccgtcgggtc atagtactgc tccaggtagg
     1441 ggtgagccag cgcttcctcc actgtgatcc gtttattggg gttaaaggtt aacatccggt
     1501 ccagcaggtc aagggctttg gagtctgact tggggaaaag cttggcccaa gccaccttgg
     1561 tcttggaggg cagagactgt aggtagtttc gggccttcat gttgatgata caattcaggt
     1621 cctcctggga tggggagccc aggatgccca gaatgtggtt gagctgatcc aggtagtgct
     1681 tgccagggaa gatgggccgg ttagagagca tctcagccag aatgcagccc acagaccaga
     1741 tgtcgatgga cttggtatag cccttggagt tcagcatgat ctctggggcc cggtaccagc
     1801 gcgtagccac atactccgtc aggaagccgg tgtggtcatg ctcaggatcg gcaatccggg
     1861 ccaggccgaa atcacaaatc ttaaggtcgc aggtggtgtt gatgagcagg ttggagggct
     1921 ttagatctcg gtggagcacg ttggcggagt ggatgtactt gaggccccgc aggatctggt
     1981 agaggaagta gcagatatgg tcattgctca gctgctggct tttcagcaac ttgtacaggt
     2041 cagtctccat caggtcctgc acaatgtaga catctctcat ggcttccagg gtggacgccc
     2101 gcagaatgtc tcggatgccg atgacattct catggcggaa gcgcagcagg atctggatct
     2161 cccggagcgt gcgctggcag taggtctgat gttcgaaggg gctgatcttc ttgatggcca
     2221 cgcgagtctt gcgcacgtgg tcataggccg agctgaccat gccgtacgcg ccctcgccga
     2281 tgtactgcaa ctgcgtgtag cgcgggccca cgtcgaacgg ctgccccttc accatctcca
     2341 cctcccccgg gacccccggg ccgaccccct cggttctacg gggctccccg cccccgcccc
     2401 cctgagccgc cgccgccgcc attgaattcg cggccgcaag cttgtcatcg tcatccttgt
     2461 aatcgatgtc atgatcttta taatcaccgt catggtcttt gtagtccatg gttaattctg
     2521 acggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     2581 ttgcgtcaac ggggcggggt tattacgaca ttttggaaag tcccgttgat tttggtgcca
     2641 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     2701 ctatccacgc ccattggtgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     2761 tacgtagatg tactgccaag taggaaagtc ccgtaaggtc atgtactggg cataatgcca
     2821 ggcgggccat ttaccgtcat tgacgtcaat agggggcgga cttggcatat gatacacttg
     2881 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     2941 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     3001 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     3061 gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca
     3121 acatggcggt catattggac atgagccaat ataaatgtac atattatgat atagatacaa
     3181 cgtatgcaat ggccaatagc caatattgat ttatgctata taaccaatga ctaatatggc
     3241 taattgccaa tattgattca atgtatagat cagcttggca ctggccgtcg ttttacaacg
     3301 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     3361 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     3421 cctgaatggc gaatggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3481 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3541 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3601 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3661 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3721 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3781 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3841 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3901 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3961 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4021 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4081 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4141 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4201 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4261 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4321 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4381 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4441 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4501 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4561 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4621 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4681 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4741 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4801 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4861 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4921 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4981 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5041 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5101 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5161 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5221 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5281 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5341 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5401 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5461 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5521 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5581 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5641 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5701 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5761 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5821 gaag
//
