LOCUS       sequence-267418-        8047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Red tagged macrophages.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8047)
  AUTHORS   Walton EM, Cronan MR, Beerman RW, Tobin DM
  TITLE     The Macrophage-Specific Promoter mfap4 Allows Live, Long-Term 
            Analysis of Macrophage Behavior during Mycobacterial Infection in 
            Zebrafish.
  JOURNAL   PLoS One. 2015 Oct 7;10(10):e0138949. doi: 
            10.1371/journal.pone.0138949. eCollection 2015.
  PUBMED    26445458
REFERENCE   2  (bases 1 to 8047)
  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: "PLoS One.";
            date: "2015-10-7"; volume: "10(10):e0138949. doi"; pages: " 
            10.1371/journal.pone.0138949. eCollection 2015"
COMMENT     Sequence Label: pDEST mfap4:tdTomato
FEATURES             Location/Qualifiers
     source          1..8047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..294
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    314..334
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        336..354
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1984..2000)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1984..2000)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2037..2055)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2055..2079
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2085..3515
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    complement(3520..3544)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3546..3680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3743..3763)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(3771..3787)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3803..3937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(4681..4868,4869..4880)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        complement(4893..4911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4918..4934)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4927..4949)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5075..5530
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5162..5181)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5372..5393
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5608..5712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5713..5781,5782..6573)
                     /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(5931..5950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6744..7332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7233..7252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7486..7503
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7620..7641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7656..7661,7662..7679,7680..7686)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7694..7710
                     /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     7699..7721
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7718..7734
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7752..7770
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7752..7769
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    join(7821..7832,7833..7970)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
ORIGIN
        1 cccaaagccc ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc
       61 ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga
      121 cagcccgggc acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc
      181 tttgagcctg cagacacctg gggggatacg gggaaaaagc tttaggctga aagagagatt
      241 tagaatgaca ggcgcgccaa ctccaatcaa ccgacgtcag gaaacagcta tgaccatgat
      301 tacgccaagc tatcaacttt gtatagaaaa gttggtaata cgactcacta tagggcgaat
      361 tgggtaccgg gccccccctc gaggcgtttc ttggtacagc tggaaaggga ataacgcggt
      421 cagtatgaaa accatcagca tcaagatcaa acaagtgcct tagaaacagg gcttttaaaa
      481 ggaaacctct tctcttttaa atgtgtatct gaaaaatcga gtctgaaatg tatggctgtt
      541 ggtgtaatga catttttctc tttgttttca ctttaaagct ttaataaact cctgtatgca
      601 gatttatgag attgtgtcct gctattcttt gaaacaaaag ctcattttcc attcaaataa
      661 tgggaactgg attaatgtac aaaactgtaa aagggcataa agcatttgtg gataaagcag
      721 agtttccatc taatgagaca aagacaacac acaataaatg caacgctgtg gcttttgaag
      781 gtgagagatg ctctttggga tgctcaagac agttctggag gtaataataa taataataat
      841 actgaaccac tgtgatgaca gactttaagg cattttacaa taagcaaaac aacatttcag
      901 atgttttaca gtgtgctgga attaaatgct gcaaagtcca ttgatgcgtc ccatcacgca
      961 cacttttttc tgatcatcat ctgttcaaat aaatcacatt acatatgctg cagctttctg
     1021 gattgtctgt agaggtttat gggtgcaagg caggagtcca gctagaagag cattgcaata
     1081 gtccagcttt ttggactaaa tgttgtgcag tatgatctct taagaagggt ctgatgttaa
     1141 agagtgcaaa caatttttca ctgagggatt cacacatgca gatatgtctg ataatcagat
     1201 catccatata aattagaaat attaagaaat cgagtggatt tacagccaga ttcttagtgt
     1261 tcgctcttaa cttgcgccat ttgattagct ctgtcacttc aggtattctt gtgtatgctg
     1321 aatcagtcaa tgttaaccca aagagaacac tcgactggtt cttgtgttag agaatttaga
     1381 gtcttgtaaa tgcagactcc aggttaacat tgcacttatc tctgcattgt gtccttttac
     1441 tcctcaacca agcttttaac caaccaatgt tttatagttg ttttgaaaaa agtcattagt
     1501 gttgtgcagg ttgaatcgtg tgtttgttat cttctgcaga aaagggttat ttggtggggt
     1561 tatcataaca tttgtctgcg tttgcagatt tggcagaaaa cagaaccgag aactgctgtc
     1621 ctttcactct tcggttgtga aaacagattg ttgggttttt gaggattgcc tcaagcaggt
     1681 tttgtcacag agcacgagac tcagcatcag cgatttaaag gagcgtttga gttatttcca
     1741 gctgaacaca cacggtgagt gacttattct gtagagcgag attatacgaa gaaataacct
     1801 gcaacaataa gaaggtgtga gtgaagctct tttacatctc ttgtagataa actgagctgt
     1861 tgaggagaga gtgagaagat tgctgtaagt tctgtggctg ttttattcct tgtgacgatc
     1921 acatttacac acgccatttt gtctcgtatt ggtgatttct ttcttcatga ctttagatcg
     1981 tgggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     2041 tagtgagggt taattcaagt ttgtacaaaa aagcaggctg gaccatggtg agcaagggcg
     2101 aggaggtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc
     2161 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     2221 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt
     2281 tcatgtacgg ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc
     2341 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg
     2401 tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc
     2461 gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     2521 cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc
     2581 tgaagctgaa ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga
     2641 agcccgtgca actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca
     2701 acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc
     2761 tggggcatgg caccggcagc accggcagcg gcagctccgg caccgcctcc tccgaggaca
     2821 acaacatggc cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga
     2881 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     2941 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     3001 cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca
     3061 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     3121 gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga
     3181 agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct
     3241 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc
     3301 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg
     3361 ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct
     3421 cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc
     3481 tgttcctgta cggcatggac gagctctaca agtagatcta cccagctttc ttgtacaaag
     3541 tgggggatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3601 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3661 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3721 gggaggtgtg ggaggttttt tccaacttta ttatacatag ttgataattc actggccgtc
     3781 gttttacggt accatcgatg atgatccaga catgataaga tacattgatg agtttggaca
     3841 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3901 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3961 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     4021 atgtggtatg gctgattatg atcctctaga tcgtgcatgc gtaccgagct cggatccccc
     4081 gggctgcagg aattctgtcc taaaacagtc tttgggcttt ctttgggggg ccggggggga
     4141 tttggagtct tttaatgatt gcttttattt tcttatatat tacaggtcta tttaaattgt
     4201 taatatgatt ttcatttaac tttggtaagt gatacctatc aagaaaattg tcatttccaa
     4261 ttttatgaag tacaggtttt tgacataatc gaattcctgc agcccggggg atccactagt
     4321 aacggccgcc agtgtgctgg aattcggctt ttagaggcag aaggagagtg aagtcaaagg
     4381 tctgcaactt cctctttgcc acataacaag aaagttgctt atatccttct ccaaggtcat
     4441 agttcctgcc agctgacttt gtcaagacag tgatgtctgt gttccagcag ttgttctgag
     4501 tatccttttc attatccact gtcctttctt cttaaattcc acccccaaca ttgtaaatag
     4561 cttctttctt aaactctgtt caaagaacca gcttgagtgt gtaagccgaa ttctgcagat
     4621 atccatcaca ctggccgcgg ttagatctat taccctgtta tccctaggga aaatagaatg
     4681 aagtgatctc caaaaaataa gtactttttg actgtaaata aaattgtaag gagtaaaaag
     4741 tacttttttt tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt tttaattgta
     4801 ctcaagtaaa gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa gtaaaattac
     4861 tcaagtactt tacacctctg ggcccaattc gccctatagt gagtcgtatt acaattcact
     4921 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     4981 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5041 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     5101 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5161 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5221 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5281 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5341 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5401 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5461 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5521 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5581 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5641 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5701 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5761 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5821 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5881 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5941 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6001 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6061 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6121 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6181 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6241 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6301 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6361 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6421 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6481 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6541 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6601 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6661 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6721 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6841 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6901 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6961 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7021 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7081 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7141 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7201 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7261 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7321 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7381 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7441 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7501 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7561 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7621 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7681 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7741 tacgccaagc tatttaggtg acactataga atactcaagc tatgcatcca acgcgttggg
     7801 agctctccca tatggtcgag cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa
     7861 agtacaagta cttagggaaa attttactca attaaaagta aaagtatctg gctagaatct
     7921 tacttgagta aaagtaaaaa agtactccat taaaattgta cttgagtatt aaggaagtag
     7981 ggataacagg gtaatctcga gcgaagttat gagttgcgcg cctgggagct cacggggaca
     8041 gcccccc
//
