LOCUS       sequence-157182-        4791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 516, Emission:
            529.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4791)
  AUTHORS   Griesbeck O, Baird GS, Campbell RE, Zacharias DA, Tsien RY
  TITLE     Reducing the environmental sensitivity of yellow fluorescent 
            protein. Mechanism and applications.
  JOURNAL   J Biol Chem. 2001 Aug 3;276(31):29188-94. Epub 2001 May 31.
  PUBMED    11387331
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: "J Biol 
            Chem."; date: "2001-08-3"; volume: "276(31)"; pages: "29188-94. Epub
            2001 May 31"
COMMENT     Sequence Label: mCitrine-pBAD
FEATURES             Location/Qualifiers
     source          1..4791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..145
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(71..90)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     231..253
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(409..1287)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1314..1598
                     /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     1521..1540
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1644..1661
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1665..1697
                     /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(1701..1709,1710..1724)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(1740..1742,1743..1745,1746..2459)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1785..1806)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2046..2065)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2393..2414
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2505..2522)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2675..2761
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2853..2880
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2899..2990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2991..3059,3060..3851)
                     /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(3209..3228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4022..4610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4511..4530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4764..4781
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
       61 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
      121 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
      181 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
      241 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg
      301 cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
      361 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
      421 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
      481 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
      541 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
      601 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
      661 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
      721 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
      781 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
      841 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
      901 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
      961 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     1021 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     1081 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     1141 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     1201 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     1261 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac
     1321 caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct
     1381 aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat
     1441 gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt
     1501 gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag cggatcctac
     1561 ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttg ggctaacagg
     1621 aggaattaac catggggggt tctcatcatc atcatcatca tggtatggct agcatgactg
     1681 gtggacagca aatgggtcgg gatctgtacg acgatgacga taaggatccg agctcgagca
     1741 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1801 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1861 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1921 tcgtgaccac cttcggctac ggcctgatgt gcttcgcccg ctaccccgac cacatgaagc
     1981 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2041 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2101 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2161 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2221 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2281 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2341 acctgagcta ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2401 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
     2461 aattcgaagc ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
     2521 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc
     2581 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
     2641 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa
     2701 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     2761 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     2821 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
     2881 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2941 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3001 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3061 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3121 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3181 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     3241 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3301 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3361 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3421 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3481 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3541 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3601 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3661 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3721 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3781 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3841 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3901 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3961 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4141 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
