LOCUS       sequence-361807-        4775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pH-sensitive RFP pHlorina for detection of efferocytosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4775)
  AUTHORS   Raymond MH, Davidson AJ, Shen Y, Tudor DR, Lucas CD, Morioka S, 
            Perry JSA, Krapivkina J, Perrais D, Schumacher LJ, Campbell RE, Wood
            W, Ravichandran KS
  TITLE     Live cell tracking of macrophage efferocytosis during Drosophila 
            embryo development in vivo.
  JOURNAL   Science. 2022 Mar 11;375(6585):1182-1187. doi: 
            10.1126/science.abl4430. Epub 2022 Mar 10.
  PUBMED    35271315
REFERENCE   2  (bases 1 to 4775)
  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"; doi: 
            "10.1126/science.abl4430"; journalName: "Science"; date: 
            "2022-03-11- 11"; volume: "375"; issue: "6585"; pages: "1182-1187"
COMMENT     Sequence Label: pBAD-pHlorina
FEATURES             Location/Qualifiers
     source          1..4775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(147..1025)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1052..1336
                     /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     1259..1278
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1382..1399
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1403..1435
                     /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(1439..1447,1448..1462)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1478..2188
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKQSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGYEASEERMYPEDGALK
                     SECKERLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERT
                     EGRHSTGGMDELYK"
     primer_bind     1891..1910
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2227..2244)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2397..2483
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2575..2602
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2621..2712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2713..2781,2782..3573)
                     /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(2931..2950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3744..4332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4233..4252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4486..4503
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4518..4658
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4584..4603)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4744..4766
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcatcaccga aacgcgcgag gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca
       61 taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag
      121 ccgtcaattg tctgattcgt taccaattat gacaacttga cggctacatc attcactttt
      181 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
      241 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
      301 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
      361 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
      421 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
      481 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
      541 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
      601 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
      661 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
      721 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg
      781 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
      841 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
      901 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
      961 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
     1021 gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga
     1081 cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt
     1141 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt
     1201 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat
     1261 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc
     1321 tctactgttt ctccataccc gttttttggg ctaacaggag gaattaacca tggggggttc
     1381 tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga
     1441 tctgtacgac gatgacgata aggatccgag ctcgagaatg gtgagcaagg gcgaggagaa
     1501 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     1561 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg cctttcagac
     1621 cgctaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     1681 tcagttcatg tacggctcca aggtttacat taagcaccca gccgacatcc ccgactactt
     1741 caagcagtcc ttccccgagg gcttcaggtg ggagcgcgtg atgaacttcg aggacggcgg
     1801 cattattcac gttaaccagg actcctccct gcaggacggc gtgttcatct acaaggtgaa
     1861 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggcta
     1921 tgaggcctcc gaggagcgga tgtaccccga ggacggcgcc ctgaagagcg agtgtaagga
     1981 gaggctgaag ctgaaggacg gcggccacta cgccgccgag gtcaagacca cctacaaggc
     2041 caagaagccc gtgcagctgc ccggcgccta catcgtcgac atcaagttgg acatcgtgtc
     2101 ccacaacgag gactacacca tcgtggaaca gtacgaacgc actgagggcc gccactccac
     2161 cggcggcatg gacgagctgt acaagtaaaa gcttggctgt tttggcggat gagagaagat
     2221 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
     2281 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
     2341 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
     2401 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     2461 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
     2521 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
     2581 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata
     2641 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2701 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2761 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2821 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2881 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2941 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3001 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3061 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3121 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3181 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3241 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3301 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3361 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3421 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3481 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3541 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3601 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3661 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3721 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3781 caaacaaaac aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3841 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3901 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3961 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4021 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4081 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4141 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4201 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4261 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4321 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4381 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4441 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4501 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4561 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     4621 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     4681 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4741 tctccgggag ctgcatgtgt cagaggtttt caccg
//
