LOCUS       sequence-418161-        2973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU1-A backbone with kanamycin resistance and pUC ORI. amilCP 
            dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2973)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 2973)
  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            "unpublished"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_UK_amilCP
FEATURES             Location/Qualifiers
     source          1..2973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(150..944)
                     /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(261..280)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     871..890
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1176..1194
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1198..1241)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1264..1298
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1307..1318
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1991..2062
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2085..2179)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2214..2231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2385..2973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2465..2484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt gaacgaaaac tcacgttaaa
       61 aagcaagcaa agaaaaaagg cgacagatta ctctgtcgcc ttttttcctg actcataacc
      121 aagccaattc tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
      181 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
      241 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
      301 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
      361 ttcggcaagc aggcatcgcc gtgggtcacg acgagatcct cgccgtcggg catacgcgcc
      421 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gatcttcgtc cagatcatcc
      481 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
      541 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
      601 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
      661 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc cgcgcaagga
      721 acgcccgtcg tkgccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca
      781 ccggacaggt cggtcttgac aaaaagaacc gggcgaccct gcgctgacag ccggaacacg
      841 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
      901 caagccgccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
      961 cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc
     1021 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc
     1081 ggcacccctt gtattactgt ttatgtaagc agacagtttt attgttgcca cctgacgtct
     1141 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt
     1201 tcagataaaa aaaatcctta gctttcgcta aggatgattt ccgtctcaat ctctatagag
     1261 acctttacag ctagctcagt cctaggtatt atgctagcta ctagagaaag aggagaaata
     1321 ctagatgagt gtgatcgcta aacaaatgac ctacaaggtt tatatgtcag gcacggtcaa
     1381 tggacactac tttgaggtcg aaggcgatgg aaaaggtaag ccctacgagg gggagcagac
     1441 ggtaaagctc actgtcacca agggcggacc tctgccattt gcttgggata ttttatcacc
     1501 acagtgtcag tacggaagca taccattcac caagtaccct gaagatatcc ctgactatgt
     1561 aaagcagtca ttcccggagg gctatacatg ggagaggatc atgaactttg aagatggtgc
     1621 agtgtgtact gtcagcaatg attccagcat ccaaggcaac tgtttcatct accatgtcaa
     1681 gttctctggt ttgaactttc ctcccaatgg acctgtcatg cagaagaaaa cacagggctg
     1741 ggaacccaac actgagcgtc tgtttgcacg agatggaatg ctgctaggaa acaactttat
     1801 ggctctgaag ttagaaggag gcggtcacta tttgtgtgaa tttaaaacta cttacaaggc
     1861 aaagaagcct gtgaagatgc cagggtatca ctatgttgac cgcaaactgg atgtaaccaa
     1921 tcacaacaag gattacactt cggttgagca gtgtgaaatt tccattgcac gcaaacctgt
     1981 ggtcgcctaa caaataaaac gaaaggatca gtcgaaagac tgggcctttc gttttatctg
     2041 ttgtttgtcg gtgaacgctc tcggtctcat gtttgccaga gacgattctc accaataaaa
     2101 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     2161 ggatctatca acaggagtcc tgcggtttcc gtgtttcgta aagtctgggc ttcctcgctc
     2221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2581 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2821 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2941 cagcagatta cgcgcagaaa aaaaggatct caa
//
