LOCUS       sequence-390615-        4436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector to restore L1 destination via Esp3I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4436)
  AUTHORS   Chamness JC, Kumar J, Cruz AJ, Rhuby E, Holum MJ, Cody JP, Tibebu R,
            Gamo ME, Starker CG, Zhang F, Voytas DF
  TITLE     An extensible vector toolkit and parts library for advanced 
            engineering of plant genomes.
  JOURNAL   Plant Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312.
  PUBMED    36896468
REFERENCE   2  (bases 1 to 4436)
  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: "Plant 
            Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312."
COMMENT     Sequence Label: pJMC-L1-Donor-Esp3I
FEATURES             Location/Qualifiers
     source          1..4436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    133..154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(169..174,175..192,193..199)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     promoter        join(681..686,687..704,705..711)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             765..1445
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             1765..2070
                     /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     1949..1968
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(2143..2162)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2144..2162)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2468..2486)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2554..2658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2659..2727,2728..3519)
                     /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(2877..2896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3690..4278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4179..4198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(4432..4436,1..13)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
       61 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcaa ttggcagtga
      121 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
      181 gcttccggct cgtataatgt gtggaattat gagcggataa taatttcaca caggaggttt
      241 aaactttaaa catgtcaaaa gagacatctt ttgttaagaa tgctgaggaa cttgcaaagc
      301 aaaaaatgga tgctattaac cctgaacttt cttcaaaatt taaattttta ataaaattcc
      361 tgtctcagtt tcctgaagct tgctctaaac ctcgttcaaa aaaaatgcag aataaagttg
      421 gtcaagagga acatattgaa tatttagctc gtagttttca tgagagtcga ttgccaagaa
      481 aacccacgcc acctacaacg gttcctgatg aggtggttag catagttctt aatataagtt
      541 ttaatataca gcctgaaaat cttgagagaa taaaagaaga acatcgattt tccatggcag
      601 ctgagaatat tgtaggagat cttctagaaa gattttcgtc tctaccggga gtgagaccgg
      661 ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat gtgtggattt
      721 tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
      781 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
      841 agtcagtcgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
      901 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
      961 tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg
     1021 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     1081 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     1141 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgttt
     1201 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     1261 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     1321 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     1381 atgaattaca acagtactgc gatgagtggc aggcggggcg taatctagag gatccggctt
     1441 actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata
     1501 tactgatatg tatacccgaa gtatgtcaaa aggaggtatg ctatgaagca gcgtattaca
     1561 gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg
     1621 gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag
     1681 cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg
     1741 ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc
     1801 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg
     1861 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
     1921 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
     1981 gtttccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
     2041 gccattaacc tgatgttctg gggaatataa ggtctcacgc tagtgagaga cgaaaatctt
     2101 gctgaaaaac tcgagccatc cggaagatct ggcggccgct ctccctatag tgagtcgtat
     2161 tacgccggat ggatatggtg ttcaggcaca agtgttaaag cagttgattt tattcactat
     2221 gatgaaaaaa acaatgaatg gaacctgctc caagttaaaa atagagataa taccgaaaac
     2281 tcatcgagta gtaagattag agataataca acaataaaaa aatggtttag aacttactca
     2341 cagcgtgatg ctactaattg ggacaatttt ccagatgaag tatcatctaa gaatttaaat
     2401 gaagaagact tcagagcttt tgttaaaaat tatttggcaa aaataatata attcggctgc
     2461 aggggcggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2521 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2581 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2641 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2701 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2761 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2821 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2881 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     2941 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3001 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3061 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3121 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3181 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3241 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3301 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3361 gccggtgagc gtgggtcacg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3421 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3481 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3541 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3601 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3661 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3721 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3781 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3841 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3901 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3961 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4021 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4081 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4141 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4201 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4261 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4321 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4381 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcga
//
