LOCUS       sequence-343623-        3778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EBFP2blue fluorescent protein CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3778)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 3778)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pEBFP2
FEATURES             Location/Qualifiers
     source          1..3778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        652..670
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     652..669
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1527..1536
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1531..1540
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1570..1589)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1571..1589)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1596..1613)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1596..1612)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1605..1627)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1753..2208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1840..1859)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2050..2071
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2286..2390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2391..2459,2460..3251)
                     /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(2609..2628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3422..3778,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tatttaggtg
      661 acactataga atactcaagc tatgcatcca acgcgttggg agctctccca tatggtcgac
      721 ctgcaggcgg ccgcgaattc actagtgatt gggcgatgag tggtctcgaa gcttacttgt
      781 acagctcatc catgcccagg gtgatgccgg cggcggtgcg gaactccagc agcaccatgt
      841 gatcgcgctt ctcgttggga tccttggaca gcacgctctg ggtgctcagg tagtggctat
      901 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtga tcggccagct
      961 gcactgagcc atcctccacg ttgtggcgga tcttgaagtt gaccttgatg ccgttcttct
     1021 gcttcacggc catgatgtag atgttgtggc tgttgaagtt gtactccagc ttgtggccca
     1081 ggatgttgcc atcctccttg aaatccacgc ccttcagctc gatgcggttc accagggtat
     1141 cgccctcgaa cttcacctcg gcgcgggtct tgtaggtgcc gtcgtcattg aagaaaatgg
     1201 tgcgctcctg cacgtagccc tcgggcatgg cgctcttgaa gaaatcgtgc tgcttcatgt
     1261 gatcggggta gcgggcgaag cactgcacgc cgtggctcag ggtggtcacc agggtgggcc
     1321 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgttggtgg
     1381 cgtcgccctc gccctcgccg cgcacgctga acttgtggcc gttcacgtcg ccatccagct
     1441 ccaccaggat gggcaccacg ccggtgaaca gctcctcgcc cttggacacc attcgagacc
     1501 actcatcgcc caatcgaatt cccgcggccg ccatggcggc cgggagcatg cgacgtcggg
     1561 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     1621 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     1681 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     1741 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1801 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1861 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1921 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     1981 cacgtagtgg gccagcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2041 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2101 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2161 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     2221 ttttctcctt acgcatctgt gcggtatttc acaccgcatc aggtggcact tttcggggaa
     2281 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2341 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2401 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2461 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2521 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2581 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2641 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2701 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2761 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2821 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2881 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2941 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3001 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3061 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtcg cgcggtatca
     3121 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3181 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3241 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3301 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3361 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3421 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3481 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3541 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3601 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3661 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3721 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaac
//
