LOCUS       sequence-418176-        4851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-a backbone with kanamycin resistance, pSC101-var1 ORI, and 
            Axe-Txe plasmid stability cassette. amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1570 to 1604)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4851)
  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            "unpublished"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_Sv1KA_amilCP
FEATURES             Location/Qualifiers
     source          1..4851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(536..1330)
                     /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(647..666)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1257..1276
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     terminator      complement(1504..1547)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1570..1604
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             1613..1624
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2297..2368
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3607..4557)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFVIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4605..4827
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa
       61 aggcttaagt agcaccctcg caagctcggt tgcggccgca atcgggcaaa tcgctgaata
      121 ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc
      181 gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg
      241 attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt
      301 tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc
      361 tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa
      421 tgcacccagt aaggcagcgg tatcatcaac aagcaaagaa aaaaggcgac agattactct
      481 gtcgcctttt ttcctgactc ataaccaagc caattctcga accccagagt cccgctcaga
      541 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
      601 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
      661 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
      721 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccgtgg gtcacgacga
      781 gatcctcgcc gtcgggcata cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
      841 cctgatgatc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
      901 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
      961 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     1021 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     1081 caacgtcgag cacagccgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     1141 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     1201 gaccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     1261 agtcatagcc gaatagcctc tccacccaag ccgccggaga acctgcgtgc aatccatctt
     1321 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc
     1381 atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac
     1441 cagagggcgc cccagctggc aattccggca ccccttgtat tactgtttat gtaagcagac
     1501 agtatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttcggt ctcactatat
     1561 ctagagacgt tgacggctag ctcagtccta ggtacagtgc tagctactag agaaagagga
     1621 gaaatactag atgagtgtga tcgctaaaca aatgacctac aaggtttata tgtcaggcac
     1681 ggtcaatgga cactactttg aggtcgaagg cgatggaaaa ggtaagccct acgaggggga
     1741 gcagacggta aagctcactg tcaccaaggg cggacctctg ccatttgctt gggatatttt
     1801 atcaccacag tgtcagtacg gaagcatacc attcaccaag taccctgaag atatccctga
     1861 ctatgtaaag cagtcattcc cggagggcta tacatgggag aggatcatga actttgaaga
     1921 tggtgcagtg tgtactgtca gcaatgattc cagcatccaa ggcaactgtt tcatctacca
     1981 tgtcaagttc tctggtttga actttcctcc caatggacct gtcatgcaga agaaaacaca
     2041 gggctgggaa cccaacactg agcgtctgtt tgcacgagat ggaatgctgc taggaaacaa
     2101 ctttatggct ctgaagttag aaggaggcgg tcactatttg tgtgaattta aaactactta
     2161 caaggcaaag aagcctgtga agatgccagg gtatcactat gttgaccgca aactggatgt
     2221 aaccaatcac aacaaggatt acacttcggt tgagcagtgt gaaatttcca ttgcacgcaa
     2281 acctgtggtc gcctaacaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2341 tatctgttgt ttgtcggtga acgctctccg tctcaccggg tacagagacc aagcgtcatc
     2401 cttagtaact aacatcccac actacaatag tattataaca tatctattca acttcaaatt
     2461 gttgaaattt aagccattta tatttacttt tcctcaactt tagaaatgat tttactaacc
     2521 ttatatccgt aatacgcgta acaaacacat tcattaataa caatcaacac taagctatat
     2581 tagcttttta aatgactagg gtaaataaaa taaggatcaa aaacttttaa gtttctgacc
     2641 ctttccttac ttccgattgg ttaatagtga tcttttgcag aataaataaa tatcgtttca
     2701 ttttcaactc tatatatcag tctatgttca tctgtaattc ttctggacca ttttccagat
     2761 aaatcatgct ttaatggctc aggttttcct aatccagcaa agggggaacg atcgatatct
     2821 tttattaact tgttaatctt ttttatattg cttttgtttc cttgctcatg ccaataaaga
     2881 taatcatccc aagcatcatc agaccaagcc ttaatcatca gattcaacct cgattaagtc
     2941 atgtgtttta aatgcacctt tggagaattg ttcatctcct cgacgaattt tttccatgac
     3001 gtaattatta gaaagtgttc tcaacgtttc ttgcatagaa tcataatctc tttttgataa
     3061 tacaacaact gtatcttcta catctttact tgttacaata agtgtttcag catcctcatt
     3121 aacttgtttc atataactac gtaaattttg gcggaaattt gaataagcta ctgcttccat
     3181 tcctttcacc ccttatttat ttctataaaa caattgtaca ttatattgta caattaagca
     3241 aatgattaat attctaactt cttataatta aaagtgctat cacttttagt ttttaggata
     3301 aaaagtgacg tcactttttg agtgctggta cctcaaatcc tgctttttct tgggcaatca
     3361 ataaatctaa cgcaatacac ataatcctgg attcactttt gccgtacttc tgctttagtt
     3421 gctgcaattc tttttgtctc ttttcatcca acgtaacggg taaccgttta tttcctattg
     3481 caattggcat tagtactctc ctagtctatg gacagttttc cctttgatat gtaacggtga
     3541 acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac aagagccata
     3601 agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg
     3661 cgtgagccat gagaacgaac cattgagatc atacttactt tgcatgtcac tcaaaaattt
     3721 tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag
     3781 tccgttatgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt
     3841 tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa
     3901 aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt
     3961 gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta
     4021 gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt
     4081 gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt
     4141 gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta actggaaaag
     4201 ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata
     4261 gtttgtccac tggaaaataa caaagccttt aaccaaagga ttcctgattt ccacagttct
     4321 cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt
     4381 catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt
     4441 ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt
     4501 taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacataca
     4561 tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta gtcaatgata
     4621 attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga cctttgctgg
     4681 aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt gtgttttttt
     4741 tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa agataaaaag
     4801 aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt a
//
