LOCUS       Exported                7023 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  luciferase assay.
ACCESSION   .
VERSION     .
KEYWORDS    miR-722-Dendra pCDNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7023)
  AUTHORS   Hsu AY, Wang D, Gurol T, Zhou W, Zhu X, Lu HY, Deng Q
  TITLE     Overexpression of microRNA-722 fine-tunes neutrophilic inflammation 
            through inhibiting Rac2 in zebrafish.
  JOURNAL   Dis Model Mech. 2017 Sep 27. pii: dmm.030791. doi: 
            10.1242/dmm.030791.
  PUBMED    28954734
REFERENCE   2  (bases 1 to 7023)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7023
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        28..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     562..582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     656..675
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        656..674
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      710..719
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1683..2375
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     primer_bind     complement(2407..2424)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2413..2637
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2683..3111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2770..2789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2980..3001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3120..3140)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3125..3454
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3305..3440
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3367..3386
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3521..4315
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3575..3594)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4185..4204
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4489..4610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4526..4545)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4580..4599
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4659..4675)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4659..4675)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4672..4694)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4683..4699
                     /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(4707..4737)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4752..4773
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4890..4907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5061..5649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5141..5160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5820..6680)
                     /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     6443..6462
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6681..6785)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6860..6879)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      601 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      661 cgactcacta tagggagacc caagctggct agcgtttaaa cttaagcttg ccaccatgga
      721 tgaggaaatc gctgccctgg tcgttgacaa cggctccggt atgtgcaaag ccggttttgc
      781 tggagatgat gcccctcgtg ctgttttccc ctccattgtt ggacgaccca gacatcaggt
      841 acatttaata aaccatccta ataacatgtt aatcatattg aaccacatta aactctcatt
      901 tgtgcatttt acatctttta aaaccgattt atattcgttt ttagataatg aagtttatta
      961 aagtgttaat tcacagcttt attgcttgtt ttgagaatct gcttgcataa gtgattttta
     1021 aacttgatct gattttcgtt tgtttcgaag ttcttcagct ttgaaagtca gatcttcaat
     1081 tcacagccat gccatagttt taggatacag tataagtaga cgaattggga tgcatcgtag
     1141 attttatgct gtgtttgtta tggtttttca gtggtagaga ttttcagagt attattaggg
     1201 ttatagtcag tgtcaggttt atttatcgtc agcgtgatgg gcaaaaaaca gctggcagta
     1261 cgggctatgt cttcgtctag aagaagacaa agccaactgc tgttactctc gcccagaaat
     1321 ctgcgccaac atggacacac ccagtggagc aagccatctg attttcatgg tcagattgta
     1381 aagaagcaat tactcttgga cagtttaagc agaaaagcca ccaagtcaag tcactggtca
     1441 tatgatgaac tcgaagatct aataaaagtt aatttaagcg ttcattacaa taaattcaag
     1501 tgaatctgtt ttaagctagc ctttaacgtt tttaaattcc ttcaaacaaa ttagttcctc
     1561 tttgcagttc ctcatttctc ggtgagctgg tgctttaatc cctttacttt ttcttcctct
     1621 agggagtgat ggttggcatg ggacgaatta attcctgcag cccgggggta ccggtcgcca
     1681 ccatgaacac cccgggaatt aacctgatca aggaggacat gcgcgtgaag gtgcacatgg
     1741 agggcaacgt gaacggccac gccttcgtga tcgagggcga gggcaagggc aagccctacg
     1801 agggcaccca gaccgccaac ctgaccgtga aggagggcgc ccccctgccc ttcagctacg
     1861 acatcctgac caccgccgtg cactacggca accgggtgtt caccaagtac cccgaggaca
     1921 tccccgacta cttcaagcag agcttccccg agggctacag ctgggagcgc accatgacct
     1981 tcgaggacaa gggcatctgc accatccgca gcgacatcag cctggagggc gactgcttct
     2041 tccagaacgt gcgcttcaag ggcaccaact tcccccccaa cggccccgtg atgcagaaga
     2101 aaaccctgaa gtgggagccc agcaccgaga agctgcacgt gcgcgacggc ctgctggtgg
     2161 gcaacatcaa catggccctg ctgctggagg gcggcggcca ctacctgtgc gacttcaaga
     2221 ccacctacaa ggccaagaag gtggtgcagc tgcccgacgc ccacttcgtg gaccaccgca
     2281 tcgagatcct gggcaacgac agcgactaca acaaggtgaa gctgtacgag cacgccgtgg
     2341 cccgctacag ccccctgccc agccaggtgt ggtaatctag agggcccgtt taaacccgct
     2401 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2461 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2521 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2581 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2641 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2701 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2761 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2821 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2881 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2941 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3001 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3061 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3121 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3181 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3301 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3361 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3421 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3481 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3541 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3601 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3661 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3721 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3781 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3841 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3901 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3961 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4021 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4081 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4141 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4201 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4261 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4321 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4381 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4441 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4501 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4561 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4621 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4681 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4741 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4801 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4861 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4921 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4981 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5041 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5101 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5161 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5221 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5281 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5341 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5401 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5461 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5521 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5581 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5641 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5701 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5761 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5821 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5881 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5941 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6001 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6061 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6121 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6181 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6241 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6301 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6361 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6421 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6481 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6541 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6601 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6661 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6721 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6781 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     6841 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     6901 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     6961 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     7021 ttc
//
