LOCUS       sequence-418163-        3543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU1-A backbone with kanamycin resistance and ColE1-bom-rop ORI.
            amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3543)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3543)
  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: 
            "unpublished"
COMMENT     Sequence Label: pTU1-A_CrK_amilCP
FEATURES             Location/Qualifiers
     source          1..3543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(124..918)
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEDHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(235..254)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     845..864
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(1057..1100)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1123..1157
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1166..1177
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1850..1921
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1944..2038)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2352..2543
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2537..2559)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2643..2782
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2698..2717
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2790..2807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2961..3543,1..6))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3041..3060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtgaac gaaaactcac
       61 gttaaagcaa gcaaagaaaa aaggcgacag attactctgt cgcctttttt cctgactcat
      121 aactcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
      181 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagttctt cagcaatatc
      241 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
      301 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccgtgggt
      361 cacgacgaga tcctcgccgt cgggcatacg cgccttgagc ctggcgaaca gttcggctgg
      421 cgcgagcccc tgatgatctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
      481 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
      541 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
      601 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
      661 ttcagtgaca acgtcgagca cagccgcgca aggaacgccc gtcgtggcca gccacgatag
      721 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
      781 aaccgggcga ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
      841 ttgtgcccag tcatagccga atagcctctc cacccaagcc gccggagaac ctgcgtgcaa
      901 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc
      961 cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc
     1021 aaccttacca gagggcgccc cagctggcaa ttccggattt cagataaaaa aaatccttag
     1081 ctttcgctaa ggatgatttc cgtctcaatc tctatagaga cctttacagc tagctcagtc
     1141 ctaggtatta tgctagctac tagagaaaga ggagaaatac tagatgagtg tgatcgctaa
     1201 acaaatgacc tacaaggttt atatgtcagg cacggtcaat ggacactact ttgaggtcga
     1261 aggcgatgga aaaggtaagc cctacgaggg ggagcagacg gtaaagctca ctgtcaccaa
     1321 gggcggacct ctgccatttg cttgggatat tttatcacca cagtgtcagt acggaagcat
     1381 accattcacc aagtaccctg aagatatccc tgactatgta aagcagtcat tcccggaggg
     1441 ctatacatgg gagaggatca tgaactttga agatggtgca gtgtgtactg tcagcaatga
     1501 ttccagcatc caaggcaact gtttcatcta ccatgtcaag ttctctggtt tgaactttcc
     1561 tcccaatgga cctgtcatgc agaagaaaac acagggctgg gaacccaaca ctgagcgtct
     1621 gtttgcacga gatggaatgc tgctaggaaa caactttatg gctctgaagt tagaaggagg
     1681 cggtcactat ttgtgtgaat ttaaaactac ttacaaggca aagaagcctg tgaagatgcc
     1741 agggtatcac tatgttgacc gcaaactgga tgtaaccaat cacaacaagg attacacttc
     1801 ggttgagcag tgtgaaattt ccattgcacg caaacctgtg gtcgcctaac aaataaaacg
     1861 aaaggatcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1921 cggtctcatg tttgccagag acgattctca ccaataaaaa acgcccggcg gcaaccgagc
     1981 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcct
     2041 gcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
     2101 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     2161 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     2221 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     2281 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     2341 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     2401 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     2461 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg
     2521 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga cggtcacagc ttgtctgtaa
     2581 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2641 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     2701 catcagagca gattgtactg agagtgcacc attgcggtgt gaaataccgc acagatgcgt
     2761 aaggagaaaa taccgcatca ggccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2821 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3361 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3541 gga
//
