LOCUS       Exported                8836 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Cre coexpressed with tdTomato in macrophages.
ACCESSION   .
VERSION     .
KEYWORDS    pDEST mfap4:icre-p2a-tomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8836)
  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 8836)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8836
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     1119..1138
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1119..1137
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2767..2783)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2820..2838)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2838..2862
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2874..2894
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(3922..3946)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3963..4019
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4026..5456
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVIKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    complement(5457..5477)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    complement(5733..5753)
                     /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(5761..5777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5793..5927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5817..5836)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5871..5890
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6599..6615)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6652..6672)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6652..6670)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6691..6707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6704..6726)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6715..6731
                     /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(6739..6769)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6784..6805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6922..6939)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7093..7681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7173..7192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7852..8712)
                     /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     8475..8494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8713..8817)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 atgctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     1081 gattacgcca agctatcaac tttgtataga aaagttggta atacgactca ctatagggcg
     1141 aattgggtac cgggcccccc ctcgaggcgt ttcttggtac agctggaaag ggaataacgc
     1201 ggtcagtatg aaaaccatca gcatcaagat caaacaagtg ccttagaaac agggctttta
     1261 aaaggaaacc tcttctcttt taaatgtgta tctgaaaaat cgagtctgaa atgtatggct
     1321 gttggtgtaa tgacattttt ctctttgttt tcactttaaa gctttaataa actcctgtat
     1381 gcagatttat gagattgtgt cctgctattc tttgaaacaa aagctcattt tccattcaaa
     1441 taatgggaac tggattaatg tacaaaactg taaaagggca taaagcattt gtggataaag
     1501 cagagtttcc atctaatgag acaaagacaa cacacaataa atgcaacgct gtggcttttg
     1561 aaggtgagag atgctctttg ggatgctcaa gacagttctg gaggtaataa taataataat
     1621 aatactgaac cactgtgatg acagacttta aggcatttta caataagcaa aacaacattt
     1681 cagatgtttt acagtgtgct ggaattaaat gctgcaaagt ccattgatgc gtcccatcac
     1741 gcacactttt ttctgatcat catctgttca aataaatcac attacatatg ctgcagcttt
     1801 ctggattgtc tgtagaggtt tatgggtgca aggcaggagt ccagctagaa gagcattgca
     1861 atagtccagc tttttggact aaatgttgtg cagtatgatc tcttaagaag ggtctgatgt
     1921 taaagagtgc aaacaatttt tcactgaggg attcacacat gcagatatgt ctgataatca
     1981 gatcatccat ataaattaga aatattaaga aatcgagtgg atttacagcc agattcttag
     2041 tgttcgctct taacttgcgc catttgatta gctctgtcac ttcaggtatt cttgtgtatg
     2101 ctgaatcagt caatgttaac ccaaagagaa cactcgactg gttcttgtgt tagagaattt
     2161 agagtcttgt aaatgcagac tccaggttaa cattgcactt atctctgcat tgtgtccttt
     2221 tactcctcaa ccaagctttt aaccaaccaa tgttttatag ttgttttgaa aaaagtcatt
     2281 agtgttgtgc aggttgaatc gtgtgtttgt tatcttctgc agaaaagggt tatttggtgg
     2341 ggttatcata acatttgtct gcgtttgcag atttggcaga aaacagaacc gagaactgct
     2401 gtcctttcac tcttcggttg tgaaaacaga ttgttgggtt tttgaggatt gcctcaagca
     2461 ggttttgtca cagagcacga gactcagcat cagcgattta aaggagcgtt tgagttattt
     2521 ccagctgaac acacacggtg agtgacttat tctgtagagc gagattatac gaagaaataa
     2581 cctgcaacaa taagaaggtg tgagtgaagc tcttttacat ctcttgtaga taaactgagc
     2641 tgttgaggag agagtgagaa gattgctgta agttctgtgg ctgttttatt ccttgtgacg
     2701 atcacattta cacacgccat tttgtctcgt attggtgatt tctttcttca tgactttaga
     2761 tcgtgggatc cactagttct agagcggccg ccaccgcggt ggagctccag cttttgttcc
     2821 ctttagtgag ggttaattca agtttgtaca aaaaagcagg ctccaccatg gtgcccaaga
     2881 agaagaggaa agtctccaac ctgctgactg tgcaccaaaa cctgcctgcc ctccctgtgg
     2941 atgccacctc tgatgaagtc aggaagaacc tgatggacat gttcagggac aggcaggcct
     3001 tctctgaaca cacctggaag atgctcctgt ctgtgtgcag atcctgggct gcctggtgca
     3061 agctgaacaa caggaaatgg ttccctgctg aacctgagga tgtgagggac tacctcctgt
     3121 acctgcaagc cagaggcctg gctgtgaaga ccatccaaca gcacctgggc cagctcaaca
     3181 tgctgcacag gagatctggc ctgcctcgcc cttctgactc caatgctgtg tccctggtga
     3241 tgaggagaat cagaaaggag aatgtggatg ctggggagag agccaagcag gccctggcct
     3301 ttgaacgcac tgactttgac caagtcagat ccctgatgga gaactctgac agatgccagg
     3361 acatcaggaa cctggccttc ctgggcattg cctacaacac cctgctgcgc attgccgaaa
     3421 ttgccagaat cagagtgaag gacatctccc gcaccgatgg tgggagaatg ctgatccaca
     3481 ttggcaggac caagaccctg gtgtccacag ctggtgtgga gaaggccctg tccctggggg
     3541 ttaccaagct ggtggagaga tggatctctg tgtctggtgt ggctgatgac cccaacaact
     3601 acctgttctg ccgggtcaga aagaatggtg tggctgcccc ttctgccacc tcccaactgt
     3661 ccacccgggc cctggaaggg atctttgagg ccacccaccg cctgatctat ggtgccaagg
     3721 atgactctgg gcagagatac ctggcctggt ctggccactc tgccagagtg ggtgctgcca
     3781 gggacatggc cagggctggt gtgtccatcc ctgaaatcat gcaggctggt ggctggacca
     3841 atgtgaacat tgtgatgaac tacatcagaa acctggactc tgagactggg gccatggtga
     3901 ggctgctcga ggatggggac gacccagctt tcttgtacaa agtggttgga tccggaagcg
     3961 gagctactaa cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggacctc
     4021 tcgagatggt gatcaagggc gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca
     4081 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     4141 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     4201 gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg
     4261 acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     4321 acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc
     4381 tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga
     4441 agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga
     4501 agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     4561 agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca
     4621 agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg
     4681 agggccgcca ccacctgttc ctggggcatg gcaccggcag caccggcagc ggcagctccg
     4741 gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa agagttcatg cgcttcaagg
     4801 tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     4861 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     4921 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc
     4981 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg
     5041 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg
     5101 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa
     5161 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg
     5221 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg
     5281 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg
     5341 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
     5401 gctccgaggg ccgccaccac ctgttcctgt acggcatgga cgagctgtac aagtaacaac
     5461 tttattatac atagttgata ttctcagtat cccctgctta tgcactcacc actttttccc
     5521 attattttat tcatgttgtg ttgctaaatt caactacatt cgtattacca tgttgtttca
     5581 tttgtttttt ttttatctca gctgtcaaaa taaaagcatt gaaatacaac attgttctat
     5641 ttttcattgt ggttttgcat gcccggtaaa taaaaaaaca aggtaataaa aataagatgc
     5701 gaaatgagaa gtataattca ctggccgtcg ttcaacttta ttatacatag ttgataattc
     5761 actggccgtc gttttacggt accatcgatg atgatccaga catgataaga tacattgatg
     5821 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     5881 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     5941 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     6001 acctctacaa atgtggtatg gctgattatg atcctctaga tcagatctgc gaagatacgg
     6061 ccacgggtgc tcttgatcct gtggctgatt ttggactgtg ctgctcgcag ctgctgatga
     6121 atcacatact tcctccattt tcttccactg attgactgtt ataatttccc taatttccag
     6181 gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac gtcacttcca aaggaccaat
     6241 gaacatgtct gaccaatttc atataatgtg aaaacgattt tcataggcag aataaataac
     6301 atttaaatta aactgggcat cagcgcaatt caattggttt ggtaatagca agggaaaata
     6361 gaatgaagtg atctccaaaa aataagtact ttttgactgt aaataaaatt gtaaggagta
     6421 aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa
     6481 ttgtactcaa gtaaagtaaa aatccccaaa aataatactt aagtacagta atcaagtaaa
     6541 attactcaag tactttacac ctctggttct tgacccccta ccttcagcaa gcccagcaga
     6601 tccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg
     6661 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6721 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6781 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6841 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6901 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6961 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7021 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7081 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7141 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7201 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7261 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7321 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7381 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7441 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7501 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7561 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7681 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7741 aggcattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7801 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7861 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7921 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7981 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8041 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8101 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8161 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8221 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8281 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8341 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8401 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8461 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8521 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8581 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8641 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8701 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8761 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8821 atttccccga aaagtg
//
