LOCUS       Exported                4006 bp ds-DNA     circular SYN 28-MAR-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4006)
  AUTHORS   UNKNOWN
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Mar 28, 2023 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4006
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     primer_bind     complement(2..21)
                     /label=A1135 JK730 2nd BsaI mut -r
                     /note="color: black; sequence: GTTTTGGCGGATGAGAGAAG; added:
                     2022-08-08"
     primer_bind     22..42
                     /label=A1134 JK730 2nd BsaI mut -f
                     /note="color: black; sequence: AGCCAAGCTGtAGACCGTTTG; 
                     added: 2022-08-08"
     misc_feature    43..779
                     /label=32 Cosecutive ISce1 site
                     /note="color: #a6acb3"
     misc_feature    824..857
                     /label=attB
                     /note="color: #a6acb3"
     misc_feature    858..926
                     /label=MSC2
                     /note="color: #a6acb3"
     primer_bind     864..909
                     /label=A1105 EcoRI-BsaI-ClaI-NheI-SacI-BsaI-SalI -f
                     /note="color: black; sequence: 
                     aattcCAGTAGAGACCatcgatgctagcgagctcGGTCTCTCCAAg; added: 
                     2022-08-17"
     primer_bind     complement(868..913)
                     /label=A1106 EcoRI-BsaI-ClaI-NheI-SacI-BsaI-SalI -r
                     /note="color: black; sequence: 
                     tcgacTTGGAGAGACCgagctcgctagcatcgatGGTCTCTACTGg; added: 
                     2022-08-17"
     misc_feature    869..908
                     /label=BsaI site
                     /note="color: #a6acb3"
     misc_feature    928..966
                     /label=attP
                     /note="color: #a6acb3"
     CDS             complement(1053..1847)
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1058..1077)
                     /label=C138 pMC-r
                     /note="color: black; sequence: tcgccttcttgacgagttct; added:
                     2022-02-04"
     primer_bind     1849..1868
                     /label=A1112 pMC Bsa deletion con-f
                     /note="color: black; sequence: cgaaacgatcctcatcctgt; added:
                     2022-07-25"
     primer_bind     complement(2149..2166)
                     /label=pMC BsaI mut -r
                     /note="color: black; sequence: cgtttggcccccctcgag; added: 
                     2022-07-18"
     primer_bind     2167..2185
                     /label=pMC BsaI mut -f
                     /note="color: black; sequence: tccccatgcgagagtaggg; added: 
                     2022-07-18"
     terminator      2199..2285
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     primer_bind     complement(2246..2265)
                     /label=A423 rrnb T1 ter -f
                     /note="color: black; sequence: cgttcaccgacaaacaacag; added:
                     2022-07-25"
     terminator      2377..2404
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     rep_origin      2538..3126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    3312..3452
                     /label=bom
                     /note="basis of mobility region from pBR322"
                     /note="color: #a6acb3"
     primer_bind     3893..3912
                     /label=pMC 2nd BsaI mut con-f
                     /note="color: black; sequence: ctggctgatacgttggtcct; added:
                     2022-08-08"
ORIGIN
        1 tcttctctca tccgccaaaa cagccaagct gtagaccgtt tgacattacc ctgttatccc
       61 tagatacatt accctgttat cccagatgac ataccctgtt atccctagat gacattaccc
      121 tgttatccca gatgacatta ccctgttatc cctagataca ttaccctgtt atcccagatg
      181 acataccctg ttatccctag atgacattac cctgttatcc cagatgacat taccctgtta
      241 tccctagata cattaccctg ttatcccaga tgacataccc tgttatccct agatgacatt
      301 accctgttat cccagatgac attaccctgt tatccctaga tacattaccc tgttatccca
      361 gatgacatac cctgttatcc ctagatgaca ttaccctgtt atcccagatg acattaccct
      421 gttatcccta gatacattac cctgttatcc cagatgacat accctgttat ccctagatga
      481 cattaccctg ttatcccaga tgacattacc ctgttatccc tagatacatt accctgttat
      541 cccagatgac ataccctgtt atccctagat gacattaccc tgttatccca gatgacatta
      601 ccctgttatc cctagataca ttaccctgtt atcccagatg acataccctg ttatccctag
      661 atgacattac cctgttatcc cagatgacat taccctgtta tccctagata cattaccctg
      721 ttatcccaga tgacataccc tgttatccct agatgacatt accctgttat cccagataaa
      781 ctcaatgatg atgatgatga tggtcgagac tcagcggccg cggtgccagg gcgtgccctt
      841 gggctccccg ggcgcgacta gtgaattcca gtagagacca tcgatgctag cgagctcggt
      901 ctctccaagt cgacccatgg gggcccgccc caactggggt aacctttgag ttctctcagt
      961 tgggggtaat cagcatcatg atgtggtacc acatcatgat gctgattata agaatgcggc
     1021 cgccacactc tagtggatct cgagttaata attcagaaga actcgtcaag aaggcgatag
     1081 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     1141 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     1201 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     1261 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc
     1321 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     1381 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     1441 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     1501 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     1561 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     1621 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg
     1681 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     1741 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     1801 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     1861 catcctgtct cttgatcaga gcttgatccc ctgcgccatc agatccttgg cggcgagaaa
     1921 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     1981 tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg
     2041 caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     2101 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtgct cgaggggggc
     2161 caaacgtccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     2221 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     2281 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     2341 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     2401 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     2461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2641 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2701 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2761 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2821 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2881 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2941 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3001 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3061 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3181 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3241 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3301 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3361 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     3421 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     3481 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3541 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca
     3601 attcgcgcgc gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga
     3661 ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg
     3721 acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc
     3781 tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat
     3841 tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga
     3901 tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg
     3961 acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgat
//
