LOCUS       sequence-437162-        4946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-a backbone with kanamycin resistance, pSC101-var5 ORI, and 
            Axe-Txe plasmid stability cassette. amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4946)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4946)
  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            "unpublished"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: pTU2-a_Sv5KA_amilCP
FEATURES             Location/Qualifiers
     source          1..4946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(546..1340)
                     /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(657..676)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1267..1286
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(1514..1557)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1580..1614
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1623..1634
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2307..2378
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2401..2495)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3712..4662)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTWKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRITLENILAKYGRI"
     rep_origin      4710..4932
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
       61 aaaaccctaa aggcttaagt agcaccctcg caagctcggt tgcggccgca atcgggcaaa
      121 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga
      181 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg
      241 ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt
      301 ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca
      361 gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca
      421 gactggctaa tgcacccagt aaggcagcgg tatcatcaac aagcaaagaa aaaaggcgac
      481 agattactct gtcgcctttt ttcctgactc ataaccaagc caattctcga accccagagt
      541 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
      601 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
      661 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
      721 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccgtgg
      781 gtcacgacga gatcctcgcc gtcgggcata cgcgccttga gcctggcgaa cagttcggct
      841 ggcgcgagcc cctgatgatc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
      901 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
      961 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     1021 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     1081 gcttcagtga caacgtcgag cacagccgcg caaggaacgc ccgtcgtggc cagccacgat
     1141 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     1201 agaaccgggc gaccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     1261 tgttgtgccc agtcatagcc gaatagcctc tccacccaag ccgccggaga acctgcgtgc
     1321 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcttgat
     1381 cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc
     1441 ccaaccttac cagagggcgc cccagctggc aattccggca ccccttgtat tactgtttat
     1501 gtaagcagac agtatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttcggt
     1561 ctcactatat ctagagacgt ttacagctag ctcagtccta ggtattatgc tagctactag
     1621 agaaagagga gaaatactag atgagtgtga tcgctaaaca aatgacctac aaggtttata
     1681 tgtcaggcac ggtcaatgga cactactttg aggtcgaagg cgatggaaaa ggtaagccct
     1741 acgaggggga gcagacggta aagctcactg tcaccaaggg cggacctctg ccatttgctt
     1801 gggatatttt atcaccacag tgtcagtacg gaagcatacc attcaccaag taccctgaag
     1861 atatccctga ctatgtaaag cagtcattcc cggagggcta tacatgggag aggatcatga
     1921 actttgaaga tggtgcagtg tgtactgtca gcaatgattc aagcatccaa ggcaactgtt
     1981 tcatctacca tgtcaagttc tctggtttga actttcctcc caatggacct gtcatgcaga
     2041 agaaaacaca gggctgggaa cccaacactg agcgtctgtt tgcacgagat ggaatgctgc
     2101 taggaaacaa ctttatggct ctgaagttag aaggaggcgg tcactatttg tgtgaattta
     2161 aaactactta caaggcaaag aagcctgtga agatgccagg gtatcactat gttgaccgca
     2221 aactggatgt aaccaatcac aacaaggatt acacttcggt tgagcagtgt gaaatttcca
     2281 ttgcacgcaa acctgtggtc gcctaacaaa taaaacgaaa ggctcagtcg aaagactggg
     2341 cctttcgttt tatctgttgt ttgtcggtga acgctctccg tctcaccggg tacagagacc
     2401 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     2461 ttctgaggtc attactggat ctatcaacag gagtcaagcg tcatccttag taactaacat
     2521 cccacactac aatagtatta taacatatct attcaacttc aaattgttga aatttaagcc
     2581 atttatattt acttttcctc aactttagaa atgattttac taaccttata tccgtaatac
     2641 gcgtaacaaa cacattcatt aataacaatc aacactaagc tatattagct ttttaaatga
     2701 ctagggtaaa taaaataagg atcaaaaact tttaagtttc tgaccctttc cttacttccg
     2761 attggttaat agtgatcttt tgcagaataa ataaatatcg tttcattttc aactctatat
     2821 atcagtctat gttcatctgt aattcttctg gaccattttc cagataaatc atgctttaat
     2881 ggctcaggtt ttcctaatcc agcaaagggg gaacgatcga tatcttttat taacttgtta
     2941 atctttttta tattgctttt gtttccttgc tcatgccaat aaagataatc atcccaagca
     3001 tcatcagacc aagccttaat catcagattc aacctcgatt aagtcatgtg ttttaaatgc
     3061 acctttggag aattgttcat ctcctcgacg aattttttcc atgacgtaat tattagaaag
     3121 tgttctcaac gtttcttgca tagaatcata atctcttttt gataatacaa caactgtatc
     3181 ttctacatct ttacttgtta caataagtgt ttcagcatcc tcattaactt gtttcatata
     3241 actacgtaaa ttttggcgga aatttgaata agctactgct tccattcctt tcacccctta
     3301 tttatttcta taaaacaatt gtacattata ttgtacaatt aagcaaatga ttaatattct
     3361 aacttcttat aattaaaagt gctatcactt ttagttttta ggataaaaag tgacgtcact
     3421 ttttgagtgc tggtacctca aatcctgctt tttcttgggc aatcaataaa tctaacgcaa
     3481 tacacataat cctggattca cttttgccgt acttctgctt tagttgctgc aattcttttt
     3541 gtctcttttc atccaacgta acgggtaacc gtttatttcc tattgcaatt ggcattagta
     3601 ctctcctagt ctatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt
     3661 ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct
     3721 tccgtattta gccagtatgt tctctagtgt gattcgttgt ttttgcgtga gccatgagaa
     3781 cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt
     3841 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta
     3901 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     3961 aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     4021 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     4081 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     4141 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     4201 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     4261 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     4321 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     4381 aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     4441 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta
     4501 ttggttataa gtgaacgata ccgtccgttc tttccatgta gggttttcaa tcgtggggtt
     4561 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     4621 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     4681 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt
     4741 cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     4801 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     4861 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     4921 tgtgtataac tcactacttt agtcag
//
