LOCUS       sequence-226991-        6331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  STARR-seq vector with 35s core promoter for transient expression in 
            plant protoplasts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6331)
  AUTHORS   Ricci WA, Lu Z, Ji L, Marand AP, Ethridge CL, Murphy NG, Noshay JM, 
            Galli M, Mejia-Guerra MK, Colome-Tatche M, Johannes F, Rowley MJ, 
            Corces VG, Zhai J, Scanlon MJ, Buckler ES, Gallavotti A, Springer 
            NM, Schmitz RJ, Zhang X
  TITLE     Widespread long-range cis-regulatory elements in the maize genome.
  JOURNAL   Nat Plants. 2019 Nov 18. pii: 10.1038/s41477-019-0547-0. doi: 
            10.1038/s41477-019-0547-0.
  PUBMED    31740773
REFERENCE   2  (bases 1 to 6331)
  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: "Nat Plants.
            2019 Nov 18. pii: 10.1038/s41477-019-0547-0. doi: 
            10.1038/s41477-019-0547-0."
COMMENT     Sequence Label: pMD19_STARR-seq_plants
FEATURES             Location/Qualifiers
     source          1..6331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(481..505)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        515..560
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     primer_bind     complement(606..634)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1229..1249
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1290..1307
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1365..1383
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     protein_bind    1392..1623
                     /gene="mutant version of attP"
                     /label=attP1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2019..2324)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(2121..2140)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2644..3327)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     3242..3261
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3328..3430)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3575..3806)
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3815..3833
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(4110..4126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4110..4126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4123..4145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4134..4150
                     /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(4158..4164,4165..4182,4183..4188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4203..4224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4341..4358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4512..5100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4592..4611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5271..6062,6063..6131))
                     /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     5894..5913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6132..6236)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6304..6322
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt agaactcggt acgcgcggat
      421 cttccagaga ttaaagaatt cgcccttata tattgatatt tatatcattt tacgtttctc
      481 gttcagcttt cttgtacaaa gtggtctcga ggatgcaaga cccttcctct atataaggaa
      541 gttcatttca tttggagagg tctagaatga gatctatgag taaaggagaa gaacttttca
      601 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
      661 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
      721 ctactggaaa actacctgtt ccatggccaa ccctggtcac caccctgacc tacggcgtgc
      781 agtgcttctc ccgttaccct gatcatatga agcggcacga cttcttcaag agcgccatgc
      841 ctgagggata cgtgcaggag aggaccatct tcttcaagga cgacgggaac tacaagacac
      901 gtgctgaagt caagtttgag ggagacaccc tcgtcaacag gatcgagctt aagggaatcg
      961 atttcaagga ggacggaaac atcctcggcc acaagttgga atacaactac aactcccaca
     1021 acgtatacat catggccgac aagcaaaaga acggcatcaa agccaacttc aagacccgcc
     1081 acaacatcga agacggcggc gtgcaactcg ctgatcatta tcaacaaaat actccaattg
     1141 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
     1201 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     1261 ttacacatgg catggtagaa ctatacaaac accaccacca ccaccactaa gatatcgccc
     1321 agaattaatt ccccaactgc acagaacaaa tcgtcggcag cgtcagatgt gtataagaga
     1381 cagagctcat caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat
     1441 tgatgagcaa tgctttttta taatgccaac tttgtacaaa aaagctgaac gagaaacgta
     1501 aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata
     1561 ctgtaaaaca caacatatcc agtcactatg aatcaactac ttagatggta ttagtgacct
     1621 gtagtcgacc gacagccttc caaatgttct tcgggtgatg ctgccaactt agtcgaccga
     1681 cagccttcca aatgttcttc tcaaacggaa tcgtcgtatc cagcctactc gctattgtcc
     1741 tcaatgccgt attaaatcat aaaaagaaat aagaaaaaga ggtgcgagcc tcttttttgt
     1801 gtgacaaaat aaaaacatct acctattcat atacgctagt gtcatagtcc tgaaaatcat
     1861 ctgcatcaag aacaatttca caactcttat acttttctct tacaagtcgt tcggcttcat
     1921 ctggattttc agcctctata cttactaaac gtgataaagt ttctgtaatt tctactgtat
     1981 cgacctgcag actggctgtg tataagggag cctgacattt atattcccca gaacatcagg
     2041 ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc ttccccgata
     2101 acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc atccccgata
     2161 tgcaccaccg ggtaaagttc acgggagact ttatctgaca gcagacgtgc actggccagg
     2221 gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc cacaaacaga
     2281 cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc agcccctgtt
     2341 ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg acctcagcca tcccttcctg
     2401 attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca tggttgtgct
     2461 taccagaccg gagatattga catcatatat gccttgagca actgatagct gtcgctgtca
     2521 actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt cgggtataca
     2581 tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac tgttatctgg
     2641 cttttagtaa gccggatcca cgcggcgttt acgccccccc tgccactcat cgcagtactg
     2701 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     2761 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     2821 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     2881 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     2941 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     3001 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     3061 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga actccggatg
     3121 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     3181 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     3241 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     3301 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     3361 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     3421 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
     3481 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
     3541 aagtgcgtcg ggtgatgctg ccaacttagt cgactacagg tcactaatac catctaagta
     3601 gttgattcat agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg
     3661 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt
     3721 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     3781 tatcagtcaa aataaaatca ttatttgatg gtacctgtct cttatacaca tctccgagcc
     3841 cacgagaccc cgggtttccc tttgcttttg tgtagacctc aaaactttat cccccatctt
     3901 tgatcttatc ccttgttttt ctgctttttt cttctttctt gggttttaat ttccggactt
     3961 aacgtttgtt ttccggtttg cgagacatat tctatcggat tctcaactgt ctgatgaaat
     4021 aaatatgtaa tgttctataa gtctttcaat ttgatgggat aagcttgtta atcgtcgaac
     4081 ggcaggcgtg caaacttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4141 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4201 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4261 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4321 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4381 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4441 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4501 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4561 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4621 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4681 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4741 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4801 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4861 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4921 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4981 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5041 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5101 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5161 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5221 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5281 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5341 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5401 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5461 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5521 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5581 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5641 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5701 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5761 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5821 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5881 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5941 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6001 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6061 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6121 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6181 atgagcggat acatatttga atgtatttag aaaaataaac aaatgggggt tccgcgcaca
     6241 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     6301 aaaaataggc gtatcacgag gccctttcgt c
//
