LOCUS       sequence-235139-        6900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  AUTHORS   Hsu AY, Wang D, Liu S, Lu J, Syahirah R, Bennin DA, Huttenlocher A, 
            Umulis DM, Wan J, Deng Q
  TITLE     Phenotypical microRNA screen reveals a noncanonical role of CDK2 in 
            regulating neutrophil migration.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Aug 26. pii: 1905221116. doi: 
            10.1073/pnas.1905221116.
  PUBMED    31451657
REFERENCE   2  (bases 1 to 6900)
  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: "Proc Natl 
            Acad Sci U S A. 2019 Aug 26. pii: 1905221116. doi: 
            10.1073/pnas.1905221116."
COMMENT     Sequence Label: vector-dendra2 PCDNA
FEATURES             Location/Qualifiers
     source          1..6900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        163..542
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        543..746
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     697..717
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     791..810
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        791..809
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1689..2381
                     /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(2419..2436)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2425..2649
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2695..3123
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2782..2801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2992..3013
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3132..3152)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3137..3466
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3317..3452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3379..3398
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3533..4327
                     /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(3587..3606)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4197..4216
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4501..4622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4538..4557)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4592..4611
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4671..4687)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4671..4687)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4684..4706)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4695..4711
                     /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(4719..4725,4726..4743,4744..4749))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4764..4785
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4902..4919)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5073..5661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5153..5172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5832..6623,6624..6692))
                     /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     6455..6474
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6693..6797)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6872..6891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
       61 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      121 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      181 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      241 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      301 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      361 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      421 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      481 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      541 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      601 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      661 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      721 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      781 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt ttaaacttaa
      841 gcttgtacat ttaataaacc atcctaataa catgttaatc atattgaacc acattaaact
      901 ctcatttgtg cattttacat cttttaaaac cgatttatat tcgtttttag ataatgaagt
      961 ttattaaagt gttaattcac agctttattg cttgttttga gaatctgctt gcataagtga
     1021 tttttaaact tgatctgatt ttcgtttgtt tcgaagttct tcagctttga aagtcagatc
     1081 ttcaattcac agccatgcca tagttttagg atacagtata agtagacgaa ttgggatgca
     1141 tcgtagattt tatgctgtgt ttgttatggt ttttcagtgg tagagatttt cagagtatta
     1201 ttagggttat agtcagtgtc aggtttattt atcgtcagcg tgatgggcaa aaaacagctg
     1261 gcagtacggg ctatgtcttc gtctagaaga agacaaagcc aactgctgtt actctcgccc
     1321 agaaatctgc gccaacatgg acacacccag tggagcaagc catctgattt tcatggtcag
     1381 attgtaaaga agcaattact cttggacagt ttaagcagaa aagccaccaa gtcaagtcac
     1441 tggtcatatg atgaactcga agatctaata aaagttaatt taagcgttca ttacaataaa
     1501 ttcaagtgaa tctgttttaa gctagccttt aacgttttta aattccttca aacaaattag
     1561 ttcctctttg cagttcctca tttctcggtg agctggtgct ttaatccctt tactttttct
     1621 tcctctaggg agtgatggtt ggcatgggac gaattaattc ctgcagcccg ggggtaccgg
     1681 tcgccaccat gaacaccccg ggaattaacc tgatcaagga ggacatgcgc gtgaaggtgc
     1741 acatggaggg caacgtgaac ggccacgcct tcgtgatcga gggcgagggc aagggcaagc
     1801 cctacgaggg cacccagacc gccaacctga ccgtgaagga gggcgccccc ctgcccttca
     1861 gctacgacat cctgaccacc gccgtgcact acggcaaccg ggtgttcacc aagtaccccg
     1921 aggacatccc cgactacttc aagcagagct tccccgaggg ctacagctgg gagcgcacca
     1981 tgaccttcga ggacaagggc atctgcacca tccgcagcga catcagcctg gagggcgact
     2041 gcttcttcca gaacgtgcgc ttcaagggca ccaacttccc ccccaacggc cccgtgatgc
     2101 agaagaaaac cctgaagtgg gagcccagca ccgagaagct gcacgtgcgc gacggcctgc
     2161 tggtgggcaa catcaacatg gccctgctgc tggagggcgg cggccactac ctgtgcgact
     2221 tcaagaccac ctacaaggcc aagaaggtgg tgcagctgcc cgacgcccac ttcgtggacc
     2281 accgcatcga gatcctgggc aacgacagcg actacaacaa ggtgaagctg tacgagcacg
     2341 ccgtggcccg ctacagcccc ctgcccagcc aggtgtggta atctagatct agagggcccg
     2401 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2461 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2521 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2581 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2641 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2701 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2761 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2821 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2881 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2941 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3001 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3061 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3121 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3241 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3301 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3361 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3421 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3481 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3541 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3601 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3661 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3721 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3781 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3841 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3901 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3961 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4021 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4081 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4141 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4201 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4261 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4321 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4381 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4441 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4501 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4561 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4621 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4681 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4741 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4801 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4861 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4921 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4981 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5881 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5941 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6001 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6061 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6121 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6181 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6241 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6301 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6361 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6421 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6481 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6541 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6601 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6661 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6721 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6781 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     6841 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
//
