LOCUS       sequence-257491-        4791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 488, Emission:
            507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4791)
  AUTHORS   Davidson Lab
  TITLE     Michael Davidson Empty Backbones
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4791)
  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            "unpublished"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: mEGFP-pBAD
FEATURES             Location/Qualifiers
     source          1..4791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    149..289
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(215..234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     375..397
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(553..1431)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1458..1742
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1665..1684
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1788..1805
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1809..1841
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1845..1853,1854..1868)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(1884..1886,1887..1889,1890..2603)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1929..1950)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2190..2209)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2537..2558
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2649..2666)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2819..2905
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2997..3024
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3043..3134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3135..3203,3204..3995)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3353..3372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4166..4754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4655..4674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
       61 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      121 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
      181 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      241 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
      301 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
      361 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
      421 cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca
      481 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
      541 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
      601 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
      661 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
      721 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
      781 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
      841 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
      901 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
      961 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     1021 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     1081 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     1141 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     1201 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     1261 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     1321 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     1381 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     1441 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     1501 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     1561 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     1621 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     1681 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc
     1741 atacccgttt tttgggctaa caggaggaat taaccatggg gggttctcat catcatcatc
     1801 atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg tacgacgatg
     1861 acgataagga tccgagctcg agcatggtga gcaagggcga ggagctgttc accggggtgg
     1921 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1981 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2041 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2101 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2161 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2221 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2281 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2341 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2401 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2461 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     2521 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2581 gcatggacga gctgtacaag taagaattcg aagcttggct gttttggcgg atgagagaag
     2641 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
     2701 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
     2761 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
     2821 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgccggt
     2881 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     2941 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
     3001 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa
     3061 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3121 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3181 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3241 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3301 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3361 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3421 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3481 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3541 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3601 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3661 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3721 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3781 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3841 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3901 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3961 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4021 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4081 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc t
//
