LOCUS       sequence-482399-        3905 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Mini circle destination vector with BsaI sites for Golden Gate 
            Assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3905)
  AUTHORS   Kang Lab
  TITLE     mini-Golden version1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3905)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pMC-Dest-BsaI
FEATURES             Location/Qualifiers
     source          1..3905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(787..804)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    825..858
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             complement(1055..1849)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1055..1849)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1166..1185)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1776..1795
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      2098..2184
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2276..2303
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      2437..3025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2926..2945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3179..3196
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3211..3351
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3277..3296)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3437..3459
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcttctctca tccgccaaaa cagccaagct gtagaccgtt tgacattacc ctgttatccc
       61 tagatacatt accctgttat cccagatgac ataccctgtt atccctagat gacattaccc
      121 tgttatccca gatgacatac cctgttatcc ctagatgaca ttaccctgtt atcccagatg
      181 acattaccct gttatcccta gatacattac cctgttatcc cagatgacat accctgttat
      241 ccctagatga cattaccctg ttatcccaga tgacattacc ctgttatccc tagatacatt
      301 accctgttat cccagatgac ataccctgtt atccctagat gacattaccc tgttatccca
      361 gatgacatta ccctgttatc cctagataca ttaccctgtt atcccagatg acataccctg
      421 ttatccctag atgacattac cctgttatcc cagatgacat taccctgtta tccctagata
      481 cattaccctg ttatcccaga tgacataccc tgttatccct agatgacatt accctgttat
      541 cccagatgac attaccctgt tatccctaga tgacattacc ctgttatccc agatgacatt
      601 accctgttat ccctagatac attaccctgt tatcccagat gacataccct gttatcccta
      661 gatgacatta ccctgttatc ccagatgaca ttaccctgtt atccctagat acattaccct
      721 gttatcccag atgacattac cctgttatcc ctagatgaca ttaccctgtt atcccagata
      781 aactcaatga tgatgatgat gatggtcgag actcagcggc cgcggtgcca gggcgtgccc
      841 ttgggctccc cgggcgcgac tagtgaattc cagtagagac catcgatgct agcgagctcg
      901 gtctctccaa gtcgacccat gggggcccgc cccaactggg gtaacctttg agttctctca
      961 gttgggggta atcagcatca tgatgtggta ccacatcatg atgctgatta taagaatgcg
     1021 gccgccacac tctagtggat ctcgagttaa taattcagaa gaactcgtca agaaggcgat
     1081 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     1141 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     1201 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     1261 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     1321 tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     1381 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     1441 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     1501 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     1561 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     1621 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca
     1681 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     1741 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     1801 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     1861 ctcatcctgt ctcttgatca gagcttgatc ccctgcgcca tcagatcctt ggcggcaaga
     1921 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     1981 attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac
     2041 tgcaagctac ctgctttctc tttgctcccc atgcgagagt agggaactgc caggcatcaa
     2101 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2161 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     2221 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     2281 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     2341 acattcaaat atgtatccgc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2401 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2461 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2521 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2581 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2641 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2701 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2761 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2821 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2881 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2941 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3001 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3061 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3121 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3181 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3241 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     3301 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3361 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3421 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3481 cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcata atgtgcctgt
     3541 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
     3601 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
     3661 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
     3721 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
     3781 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
     3841 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     3901 gcgat
//
