LOCUS       sequence-267420-        6918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyan tagged macrophages.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6918)
  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 6918)
  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:turquoise2
FEATURES             Location/Qualifiers
     source          1..6918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..47
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    67..87
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     89..108
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        89..107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1737..1753)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1790..1808)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    1808..1832
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1838..1840,1841..1843,1844..2557)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1883..1904)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2144..2163)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2491..2512
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2562..2586)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2588..2722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(2785..2805)
                     /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(2813..2829)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2845..2979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3418..3605,3606..3617)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3651..3667)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3704..3722)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3743..3759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3756..3778)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3767..3783
                     /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)."
     promoter        complement(join(3791..3797,3798..3815,3816..3821))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3836..3857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3974..3991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4145..4733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4225..4244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4904..5695,5696..5764))
                     /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     5527..5546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5765..5869)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6032..6053)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6402..6413,6414..6576)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
ORIGIN
        1 tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat gattacgcca
       61 agctatcaac tttgtataga aaagttggta atacgactca ctatagggcg aattgggtac
      121 cgggcccccc ctcgaggcgt ttcttggtac agctggaaag ggaataacgc ggtcagtatg
      181 aaaaccatca gcatcaagat caaacaagtg ccttagaaac agggctttta aaaggaaacc
      241 tcttctcttt taaatgtgta tctgaaaaat cgagtctgaa atgtatggct gttggtgtaa
      301 tgacattttt ctctttgttt tcactttaaa gctttaataa actcctgtat gcagatttat
      361 gagattgtgt cctgctattc tttgaaacaa aagctcattt tccattcaaa taatgggaac
      421 tggattaatg tacaaaactg taaaagggca taaagcattt gtggataaag cagagtttcc
      481 atctaatgag acaaagacaa cacacaataa atgcaacgct gtggcttttg aaggtgagag
      541 atgctctttg ggatgctcaa gacagttctg gaggtaataa taataataat aatactgaac
      601 cactgtgatg acagacttta aggcatttta caataagcaa aacaacattt cagatgtttt
      661 acagtgtgct ggaattaaat gctgcaaagt ccattgatgc gtcccatcac gcacactttt
      721 ttctgatcat catctgttca aataaatcac attacatatg ctgcagcttt ctggattgtc
      781 tgtagaggtt tatgggtgca aggcaggagt ccagctagaa gagcattgca atagtccagc
      841 tttttggact aaatgttgtg cagtatgatc tcttaagaag ggtctgatgt taaagagtgc
      901 aaacaatttt tcactgaggg attcacacat gcagatatgt ctgataatca gatcatccat
      961 ataaattaga aatattaaga aatcgagtgg atttacagcc agattcttag tgttcgctct
     1021 taacttgcgc catttgatta gctctgtcac ttcaggtatt cttgtgtatg ctgaatcagt
     1081 caatgttaac ccaaagagaa cactcgactg gttcttgtgt tagagaattt agagtcttgt
     1141 aaatgcagac tccaggttaa cattgcactt atctctgcat tgtgtccttt tactcctcaa
     1201 ccaagctttt aaccaaccaa tgttttatag ttgttttgaa aaaagtcatt agtgttgtgc
     1261 aggttgaatc gtgtgtttgt tatcttctgc agaaaagggt tatttggtgg ggttatcata
     1321 acatttgtct gcgtttgcag atttggcaga aaacagaacc gagaactgct gtcctttcac
     1381 tcttcggttg tgaaaacaga ttgttgggtt tttgaggatt gcctcaagca ggttttgtca
     1441 cagagcacga gactcagcat cagcgattta aaggagcgtt tgagttattt ccagctgaac
     1501 acacacggtg agtgacttat tctgtagagc gagattatac gaagaaataa cctgcaacaa
     1561 taagaaggtg tgagtgaagc tcttttacat ctcttgtaga taaactgagc tgttgaggag
     1621 agagtgagaa gattgctgta agttctgtgg ctgttttatt ccttgtgacg atcacattta
     1681 cacacgccat tttgtctcgt attggtgatt tctttcttca tgactttaga tcgtgggatc
     1741 cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     1801 ggttaattca agtttgtaca aaaaagcagg ctggaccatg gtgagcaagg gcgaggagct
     1861 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1921 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1981 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg
     2041 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2101 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2161 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2221 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actactttag
     2281 cgacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     2341 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     2401 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     2461 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2521 cgggatcact ctcggcatgg acgagctcta caagtagatc tacccagctt tcttgtacaa
     2581 agtgggggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2641 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2701 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2761 gggggaggtg tgggaggttt tttccaactt tattatacat agttgataat tcactggccg
     2821 tcgttttacg gtaccatcga tgatgatcca gacatgataa gatacattga tgagtttgga
     2881 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2941 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3001 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     3061 aaatgtggta tggctgatta tgatcctcta gatcagatct gcgaagatac ggccacgggt
     3121 gctcttgatc ctgtggctga ttttggactg tgctgctcgc agctgctgat gaatcacata
     3181 cttcctccat tttcttccac tgattgactg ttataatttc cctaatttcc aggtcaaggt
     3241 gctgtgcatt gtggtaatag atgtgacatg acgtcacttc caaaggacca atgaacatgt
     3301 ctgaccaatt tcatataatg tgaaaacgat tttcataggc agaataaata acatttaaat
     3361 taaactgggc atcagcgcaa ttcaattggt ttggtaatag caagggaaaa tagaatgaag
     3421 tgatctccaa aaaataagta ctttttgact gtaaataaaa ttgtaaggag taaaaagtac
     3481 ttttttttct aaaaaaatgt aattaagtaa aagtaaaagt attgattttt aattgtactc
     3541 aagtaaagta aaaatcccca aaaataatac ttaagtacag taatcaagta aaattactca
     3601 agtactttac acctctggtt cttgaccccc taccttcagc aagcccagca gatccactag
     3661 ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa
     3721 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3781 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3841 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3901 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3961 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4021 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4081 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4321 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4561 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taaggcattt
     4801 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4861 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4921 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4981 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5041 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5101 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5161 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5221 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5281 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5341 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5401 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5461 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5521 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5581 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5641 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5701 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5761 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5821 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5881 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5941 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     6001 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     6061 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccacgctg
     6121 gctaagaact catcagcctc cccggtccat ctacccacgt accaatgcac caattggcca
     6181 caatgacggc tactacatgg tgccattcct tcctctttat aggaatggag actacctcct
     6241 gtccaacaat gctcttggat acgagtacgc ctacctgttg gacccaggtc attgcacaac
     6301 accagaaatg ccctctgatc tgcaaaagac gtgaatatct gttcagacac ccatatccac
     6361 tctgttccac acaggtcaga ggtttgtcca ggagttcttg acagaggtgt aaaaagtact
     6421 caaaaatttt actcaagtga aagtacaagt acttagggaa aattttactc aattaaaagt
     6481 aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa aagtactcca ttaaaattgt
     6541 acttgagtat taaggaagta aaagtaaaag caagaaagaa aactagagat tcttgtttaa
     6601 gcttttaatc tcaaaaaaca ttaaatgaaa tgcatacaag gttttatcct gctttagaac
     6661 tgtttgtatt taattatcaa actataagac agacaatcta atgccagtac acgctactca
     6721 aagttgtaaa acctcagatt taacttcagt agaagctgat tctcaaaatt gttagtgtca
     6781 agcctagctc ttttggggct gaaaagcaat cctgcagtgc tgaaaagcct ctcacaggca
     6841 gccgatgcgg gaagaggtgt attagtcttg atagagaggc tgcaaatagc aggaaacgtg
     6901 agcagagact ccctggtg
//
