LOCUS       sequence-114008-        8966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression vector for MARK2 with a C-terminal eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8966)
  TITLE     Banker Lab MARK2-GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8966)
  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            "Unpublished"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: pBa-MARK2-eGFP
FEATURES             Location/Qualifiers
     source          1..8966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        418..693
                     /label=chicken beta-actin promoter
     primer_bind     1626..1648
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    1713..1737
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3913..3937)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3954..3956,3957..3959,3960..4670)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGGELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3999..4020)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4260..4279)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4607..4628
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4803..4937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4840..4859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4894..4913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4969..4985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4969..4985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4982..5004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4993..5009
                     /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(5017..5023,5024..5041,5042..5047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5062..5083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5200..5217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5371..5959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5451..5470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6130..6921,6922..6990))
                     /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     6753..6772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6991..7095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7163..7181
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7219..7241)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7341..7360
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7554..7576
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7568..7585
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7569..7585
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttagtgcatg catacccctc ggtgttattg ctgctctgtg cacgcacgct cattgtatca
       61 cttcatccca gtgcatgcac tcacactgga gcgattgctg ctcggtgcac gcacactcat
      121 tgtatcacgt cagctcagtg gctgcacgca caccggtgtt attgctgctc ggtgcgtgca
      181 tgcacatcag tgtcgctgca gctcagtgca tgcacgctca ttgcccatcg ctatccctgc
      241 ctctcctgct ggcgctcccc gggaggtgac ttcaagggga ccgcaggacc acctcggggg
      301 tggggggagg gctgcacacg cggaccccgc tccccctccc caacaaagca ctgtggaatc
      361 aaaaaggggg gaggggggat ggaggggcgc gtcacacccc cgccccacac cctcacctcg
      421 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      481 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
      541 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
      601 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      661 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt
      721 gccccgctcc gcgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      781 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
      841 cggctcgttt cttttctgtg gctgcgtgaa agccttaaag ggctccggga gggccctttg
      901 tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
      961 cggcccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1021 cgcgtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga
     1081 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1141 ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     1201 cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     1261 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     1321 aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1381 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctggcggag
     1441 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg
     1501 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     1561 tccatctcca gcctcggggc tgccgcaggg ggacggctgc cttcgggggg gacggggcag
     1621 ggcggggttc ggcttctggc gtgtgaccgg cggggtttat atcttccctt ctctgttcct
     1681 ccgcagcccc caagcttggc gcgcctataa tgccaacttt gtacaaaaaa gttggcacca
     1741 tgtccagcgc tcggaccccc ctacccacgc tgaacgagag ggacacggag cagcccacct
     1801 tgggacacct tgactccaag cccagcagta agtccaacat gattcggggc cgcaactcag
     1861 ccacctctgc tgatgagcag ccccacattg gaaactaccg gctcctcaag accattggca
     1921 agggtaattt tgccaaggtg aagttggccc gacacatcct gactgggaaa gaggtagctg
     1981 tgaagatcat tgacaagact caactgaact cctccagcct ccagaaacta ttccgcgaag
     2041 taagaataat gaaggttttg aatcatccca acatagttaa attatttgaa gtgattgaga
     2101 ctgagaaaac gctctacctt gtcatggagt acgctagtgg cggagaggta tttgattacc
     2161 tagtggctca tggcaggatg aaagaaaaag aggctcgagc caaattccgc cagatagtgt
     2221 ctgctgtgca gtactgtcac cagaagttta ttgtccatag agacttaaag gcagaaaacc
     2281 tgctcttgga tgctgatatg aacatcaaga ttgcagactt tggcttcagc aatgaattca
     2341 cctttgggaa caagctggac accttctgtg gcagtccccc ttatgctgcc ccagaactct
     2401 tccagggcaa aaaatatgat ggacccgagg tggatgtgtg gagcctagga gttatcctct
     2461 atacactggt cagcggatcc ctgccttttg atggacagaa cctcaaggag ctgcgggaac
     2521 gggtactgag gggaaaatac cgtattccat tctacatgtc cacggactgt gaaaacctgc
     2581 ttaagaaatt tctcattctt aatcccagca agagaggcac tttagagcaa atcatgaaag
     2641 atcgatggat gaatgtgggt cacgaagatg atgaactaaa gccttacgtg gagccactcc
     2701 ctgactacaa ggacccccgg cggacagagc tgatggtgtc catgggttat acacgggaag
     2761 agatccagga ctcgctggtg ggccagagat acaacgaggt gatggccacc tatctgctcc
     2821 tgggctacaa gagctccgag ctggaaggcg acaccatcac cctgaaaccc cggccttcag
     2881 ctgatctgac caatagcagc gccccatccc catcccacaa ggtacagcgc agcgtgtcgg
     2941 ccaatcccaa gcagcggcgc ttcagcgacc aggctggtcc tgccattccc acctctaatt
     3001 cttactctaa gaagactcag agtaacaacg cagaaaataa gcggcctgag gaggaccggg
     3061 agtcagggcg gaaagccagc agcacagcca aggtgcctgc cagccccctg cccggtctgg
     3121 agaggaagaa gaccacccca accccctcca cgaacagcgt cctctccacc agcacaaatc
     3181 gaagcaggaa ttccccactt ttggagcggg ccagcctcgg ccaggcctcc atccagaatg
     3241 gcaaagacag cacagccccc cagcgtgtcc ctgttgcctc cccatccgcc cacaacatca
     3301 gcagcagtgg tggagcccca gaccgaacta acttcccccg gggtgtgtcc agccgaagca
     3361 ccttccatgc tgggcagctc cgacaggtgc gggaccagca gaatttgccc tacggtgtga
     3421 ccccagcctc tccctctggc cacagccagg gccggcgggg ggcctctggg agcatcttca
     3481 gcaagttcac ctccaagttt gtacgcagga acctgaatga acctgaaagc aaagaccgag
     3541 tggagacgct cagacctcac gtggtgggca gtggcggcaa cgacaaagaa aaggaagaat
     3601 ttcgggaggc caagccccgc tccctccgct tcacgtggag tatgaagacc acgagctcca
     3661 tggagcccaa cgagatgatg cgggagatcc gcaaggtgct ggacgcgaac agctgccaga
     3721 gcgagctgca tgagaagtac atgctgctgt gcatgcacgg cacgccgggc cacgaggact
     3781 tcgtgcagtg ggagatggag gtgtgcaaac tgccgcggct ctctctcaac ggggttcgat
     3841 ttaagcggat atcgggcacc tccatggcct tcaaaaacat tgcctccaaa atagccaacg
     3901 agctgaagct tttgccaact ttcttgtaca aagttggcat tactaggcct accatggtga
     3961 gcaagggcgg ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4021 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     4081 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     4141 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     4201 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     4261 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     4321 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     4381 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     4441 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     4501 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     4561 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     4621 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag cggaccgtct
     4681 aatctagata agtaatgatc ataatcagcc atatcacatc tgtagaggtt ttacttgctt
     4741 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     4801 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4861 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4921 cttatcatgt ctggatcccc gggtaccgag ctcgaattcg taatcatggt catagctgtt
     4981 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5041 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5101 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5161 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5221 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5281 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5341 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5401 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5461 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5521 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5581 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5641 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5701 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5761 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5821 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5881 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5941 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6001 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6061 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6121 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6181 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6241 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6301 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6361 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6421 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6481 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6541 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6601 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6661 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6721 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6781 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6841 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6901 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6961 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7021 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7081 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7141 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     7201 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     7261 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     7321 tcggggttgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     7381 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     7441 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     7501 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7561 acgacgttgt aaaacgacgg ccagtgccaa gttgggatct ttgcattggc ccacggctct
     7621 caggatgggg atgctcccct tcagcacccg gttccccttg gaaactgatg gtcctggctc
     7681 tgtggcatgg cagtggcact gtgaggagcc cctaccagca gcacacagtg ggtttggcac
     7741 tgccacgctc cggatgccgc gctctgatcc aaccccataa tcaagggaac ccgaattgcc
     7801 ccatcattgc ccccaccacc cccatcctgc cgggccctca caccccacgc tgccttgtgg
     7861 tgacattccc cagcccaaac ccacggcttc atggctaccg cggggcattt cccattgccg
     7921 ccccattatc agctctgcac acctcccgct gtacccatgc ctcgtggctg cccttctttg
     7981 acgtataatc ttctaattaa tacccggcct tgtcaaagtg gagcacaaac gttaattaat
     8041 tccccagcag gcaggtaatt aacagtgtga ctcccttttt gctgcgagtg gggctgatac
     8101 agagagatgt ggcactatgg agcccacggg gtcctggcac tgggtgccca cggaggtccc
     8161 catgtgctgc agtgtcaccg cctccgaggt gacagtattg tccctgcggt gtccctgcag
     8221 ctcagctctg tccacagggc cacctccagt ttggagggga cacaatgcag ccccgatgca
     8281 acccatcctc gcagcatccc agggacaaag accccactgc aagaccgcac acagggctgg
     8341 gtcccgctcc cctaatatct acagtgcttt tgcatggccc cttaatcaat gcagttaatc
     8401 agcatgcgct catgcaccgc tctggagctg caaagcccct cgcagcgctg ctcaccaaca
     8461 ccgcgcaccg ccccggccca gcctgcagca cgcgctgcaa acaggaaaga aacaaaatat
     8521 tgcccaaatg taggcaaagg cattcggctg ccttgacctc cgccgggccg ggccctgcct
     8581 gactcagctc cttactcagc gctcgcttcc tccctccggc tgccaccgcc gcagcgcaca
     8641 ccctgacaaa gagtggccct taacgggctc tgaggtgcac ccagcagtgc actcagcagt
     8701 ccaagggccg gcctggaggt ttgcaccgct acgtgctgac attagcattg aacttggccc
     8761 tgggtagtgc tgcaggccgg gcggggtggg tgtagagagt gcagcgcgcg ttgcacccgg
     8821 tgccccttcc cctcccttgc atcccagcag gctgcacccc agcaccaggc ccgtgcatgc
     8881 atgctcctgg tgttattgca gcctggtgca tgcatgcgtc ttagtggtgc agcgctgtgc
     8941 atgcatcctc cttggtgtgt agcagc
//
