LOCUS       sequence-376795-        4374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for inducible mNeonGreen expression with 4x LexA operator 
            (LexAO).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4374)
  AUTHORS   Huang Z, Wu Y, Allen ME, Pan Y, Kyriakakis P, Lu S, Chang YJ, Wang 
            X, Chien S, Wang Y
  TITLE     Engineering light-controllable CAR T cells for cancer immunotherapy.
  JOURNAL   Sci Adv. 2020 Feb 19;6(8):eaay9209. doi: 10.1126/sciadv.aay9209. 
            eCollection 2020 Feb.
  PUBMED    32128416
REFERENCE   2  (bases 1 to 4374)
  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: "Sci Adv."; 
            date: "2020-02-19"; volume: "6(8):eaay9209. doi"; pages: " 
            10.1126/sciadv.aay9209. eCollection 2020 Feb"
COMMENT     Sequence Label: pDN-LexAOx4-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..4374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             93..803
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      348..357
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          934..999
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1129..1149
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1574..1708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1611..1630)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1665..1684
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1981..1999)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2067..2171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2172..2240,2241..3032)
                     /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(2390..2409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3203..3791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3692..3711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3945..3962
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3977..4117
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4043..4062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctcgagtccg agcggagact ctagagggta tataatggat ccccgggtac cgagctcgaa
       61 ttccagcttg gcattccggt actgttggta aaatggtgag caagggcgag gaggataaca
      121 tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact
      181 ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga
      241 agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg
      301 gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag
      361 atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg
      421 ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga
      481 ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca
      541 ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca
      601 ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc
      661 caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca
      721 agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg
      781 gcatggacga gctgtacaag taaaatgtaa ctgtattcag cgatgacgaa attcttagct
      841 attgtaatac tctagaggat ctttgtgaag gaaccttact tctgtggtgt gacataattg
      901 gacaaactac ctacagagat ttaaagctct aaggtaaata taaaattttt aagtgtataa
      961 tgtgttaaac tactgattct aattgtttgt gtattttaga ttccaaccta tggaactgat
     1021 gaatgggagc agtggtggaa tgcctttaat gaggaaaacc tgttttgctc agaagaaatg
     1081 ccatctagtg atgatgaggc tactgctgac tctcaacatt ctactcctcc aaaaaagaag
     1141 agaaaggtag aagaccccaa ggactttcct tcagaattgc taagtttttt gagtcatgct
     1201 gtgtttagta atagaactct tgcttgcttt gctatttaca ccacaaagga aaaagctgca
     1261 ctgctataca agaaaattat ggaaaaatat tctgtaacct ttataagtag gcataacagt
     1321 tataatcata acatactgtt ttttcttact ccacacaggc atagagtgtc tgctattaat
     1381 aactatgctc aaaaattgtg tacctttagc tttttaattt gtaaaggggt taataaggaa
     1441 tatttgatgt atagtgcctt gactagagat cataatcagc cataccacat ttgtagaggt
     1501 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     1561 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1621 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1681 catcaatgta tcttatcatg tctggatccc caggaagctc ctctgtgtcc tcataaaccc
     1741 taacctcctc tacttgagag gacattccaa tcataggctg cccatccacc ctctgtgtcc
     1801 tcctgttaat taggtcactt aacaaaaagg aaattgggta ggggtttttc acagaccgct
     1861 ttctaagggt aattttaaaa tatctgggaa gtcccttcca ctgctgtgtt ccagaagtgt
     1921 tggtaaacag cccacaaatg tcaacagcag aaacatacaa gctgtcagct ttgcacaagg
     1981 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     2041 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2101 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2161 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2221 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2281 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2341 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2401 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2461 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2521 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2581 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2641 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2701 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2761 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2821 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2881 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2941 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3001 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3061 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3121 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3181 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3241 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3301 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3361 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3421 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3481 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3541 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3601 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3661 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3721 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3781 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3841 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3901 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3961 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4021 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     4081 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     4141 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgg
     4201 gtaccgaatt cgcggccgca tctagactgt atataaaacc agtggttata tgtacagact
     4261 agactgtata taaaaccagt ggttatatgt acagactaga ctgtatataa aaccagtggt
     4321 tatatgtaca gactagactg tatataaaac cagtggttat atgtacagac taga
//
