LOCUS       sequence-418164-        4946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-a backbone with kanamycin resistance, pSC101-var7 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  JOURNAL   unpublished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            "unpublished"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_Sv7KA_amilCP
FEATURES             Location/Qualifiers
     source          1..4946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(551..1345)
                     /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(662..681)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1272..1291
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(1519..1562)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1585..1619
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     terminator      2312..2383
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2406..2500)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3717..4667)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTDYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLGNNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4715..4937
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag
       61 accttaaaac cctaaaggct taagtagcac cctcgcaagc tcggttgcgg ccgcaatcgg
      121 gcaaatcgct gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt
      181 cgtgacattc agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac
      241 aggcgccttt tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg
      301 ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt
      361 cagcagttcc tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc
      421 attcagactg gctaatgcac ccagtaaggc agcggtatca tcaacaagca aagaaaaaag
      481 gcgacagatt actctgtcgc cttttttcct gactcataac caagccaatt ctcgaacccc
      541 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
      601 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
      661 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
      721 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
      781 cgtgggtcac gacgagatcc tcgccgtcgg gcatacgcgc cttgagcctg gcgaacagtt
      841 cggctggcgc gagcccctga tgatcttcgt ccagatcatc ctgatcgaca agaccggctt
      901 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
      961 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     1021 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     1081 ttcccgcttc agtgacaacg tcgagcacag ccgcgcaagg aacgcccgtc gtggccagcc
     1141 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     1201 caaaaagaac cgggcgaccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     1261 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagccgcc ggagaacctg
     1321 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc
     1381 ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg
     1441 gcttcccaac cttaccagag ggcgccccag ctggcaattc cggcacccct tgtattactg
     1501 tttatgtaag cagacagtat ttcagataaa aaaaatcctt agctttcgct aaggatgatt
     1561 tcggtctcac tatatctaga gacgtttaca gctagctcag tcctaggtat tatgctagct
     1621 actagagaaa gaggataaat actagatgag tgtgatcgct aaacaaatga cctacaaggt
     1681 ttatatgtca ggcacggtca atggacacta ctttgaggtc gaaggcgatg gaaaaggtaa
     1741 gccctacgag ggggagcaga cggtaaagct cactgtcacc aagggcggac ctctgccatt
     1801 tgcttgggat attttatcac cacagtgtca gtacggaagc ataccattca ccaagtaccc
     1861 tgaagatatc cctgactatg taaagcagtc attcccggag ggctatacat gggagaggat
     1921 catgaacttt gaagatggtg cagtgtgtac tgtcagcaat gattccagca tccaaggcaa
     1981 ctgtttcatc taccatgtca agttctctgg tttgaacttt cctcccaatg gacctgtcat
     2041 gcagaagaaa acacagggct gggaacccaa cactgagcgt ctgtttgcac gagatggaat
     2101 gctgctagga aacaacttta tggctctgaa gttagaagga ggcggtcact atttgtgtga
     2161 atttaaaact acttacaagg caaagaagcc tgtgaagatg ccagggtatc actatgttga
     2221 ccgcaaactg gatgtaacca atcacaacaa ggattacact tcggttgagc agtgtgaaat
     2281 ttccattgca cgcaaacctg tggtcgccta acaaataaaa cgaaaggctc agtcgaaaga
     2341 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctccgtctca ccgggtacag
     2401 agaccattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     2461 tggagttctg aggtcattac tggatctatc aacaggagtc aagcgtcatc cttagtaact
     2521 aacatcccac actacaatag tattataaca tatctattca acttcaaatt gttgaaattt
     2581 aagccattta tatttacttt tcctcaactt tagaaatgat tttactaacc ttatatccgt
     2641 aatacgcgta acaaacacat tcattaataa caatcaacac taagctatat tagcttttta
     2701 aatgactagg gtaaataaaa taaggatcaa aaacttttaa gtttctgacc ctttccttac
     2761 ttccgattgg ttaatagtga tcttttgcag aataaataaa tatcgtttca ttttcaactc
     2821 tatatatcag tctatgttca tctgtaattc ttctggacca ttttccagat aaatcatgct
     2881 ttaatggctc aggttttcct aatccagcaa agggggaacg atcgatatct tttattaact
     2941 tgttaatctt ttttatattg cttttgtttc cttgctcatg ccaataaaga taatcatccc
     3001 aagcatcatc agaccaagcc ttaatcatca gattcaacct cgattaagtc atgtgtttta
     3061 aatgcacctt tggagaattg ttcatctcct cgacgaattt tttccatgac gtaattatta
     3121 gaaagtgttc tcaacgtttc ttgcatagaa tcataatctc tttttgataa tacaacaact
     3181 gtatcttcta catctttact tgttacaata agtgtttcag catcctcatt aacttgtttc
     3241 atataactac gtaaattttg gcggaaattt gaataagcta ctgcttccat tcctttcacc
     3301 ccttatttat ttctataaaa caattgtaca ttatattgta caattaagca aatgattaat
     3361 attctaactt cttataatta aaagtgctat cacttttagt ttttaggata aaaagtgacg
     3421 tcactttttg agtgctggta cctcaaatcc tgctttttct tgggcaatca ataaatctaa
     3481 cgcaatacac ataatcctgg attcactttt gccgtacttc tgctttagtt gctgcaattc
     3541 tttttgtctc ttttcatcca acgtaacggg taaccgttta tttcctattg caattggcat
     3601 tagtactctc ctagtctatg gacagttttc cctttgatat gtaacggtga acagttgttc
     3661 tacttttgtt tgttagtctt gatgcttcac tgatagatac aagagccata agaacctcag
     3721 atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat
     3781 gagaacgaac cattgagatc atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa
     3841 ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttatgt
     3901 aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg
     3961 ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta
     4021 tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct
     4081 ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta gttatttcca
     4141 agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc
     4201 ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa
     4261 gacttaacat gttccagatt atattttatg aattttttta actggaaaag ataaggcaat
     4321 atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata gtctgtccac
     4381 tggaaaatct caaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc
     4441 tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga
     4501 gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg
     4561 gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag
     4621 cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacataca tctcaattgg
     4681 tctaggtgat tttaatcact ataccaattg agatgggcta gtcaatgata attactagtc
     4741 cttttccttt gagttgtggg tatctgtaaa ttctgctaga cctttgctgg aaaacttgta
     4801 aattctgcta gaccctctgt aaattccgct agacctttgt gtgttttttt tgtttatatt
     4861 caagtggtta taatttatag aataaagaaa gaataaaaaa agataaaaag aatagatccc
     4921 agccctgtgt ataactcact acttta
//
