LOCUS       sequence-418185-        3910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-b backbone with kanamycin resistance and pSC101-var4 ORI. 
            amilCP dropout vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3910)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3910)
  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            "unpublished"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-b_Sv4K_amilCP
FEATURES             Location/Qualifiers
     source          1..3910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(547..1341)
                     /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(658..677)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1268..1287
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1573..1591
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1595..1638)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1661..1695
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1704..1715
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2388..2459
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2482..2576)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2677..3627)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFGIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3675..3897
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct
       61 taaaacccta aaggcttaag tagcaccctc gcaagctcgg ttgcggccgc aatcgggcaa
      121 atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg
      181 acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc
      241 gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct
      301 tctcagggcg ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc
      361 agttcctgcc ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc
      421 agactggcta atgcacccag taaggcagcg gtatcatcaa caagcaaaga aaaaaggcga
      481 cagattactc tgtcgccttt tttcctgact cataaccaag ccaattctcg aaccccagag
      541 tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
      601 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
      661 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
      721 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccgtg
      781 ggtcacgacg agatcctcgc cgtcgggcat acgcgccttg agcctggcga acagttcggc
      841 tggcgcgagc ccctgatgat cttcgtccag atcatcctga tcgacaagac cggcttccat
      901 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
      961 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     1021 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     1081 cgcttcagtg acaacgtcga gcacagccgc gcaaggaacg cccgtcgtgg ccagccacga
     1141 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     1201 aagaaccggg cgaccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     1261 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gccgccggag aacctgcgtg
     1321 caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcttga
     1381 tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt
     1441 cccaacctta ccagagggcg ccccagctgg caattccggc accccttgta ttactgttta
     1501 tgtaagcaga cagttttatt gttgccacct gacgtctaag aaaccattat tatcatgaca
     1561 ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct
     1621 ttcgctaagg atgatttcgg tctcactata tctagagacg tttacagcta gctcagtcct
     1681 aggtattatg ctagctacta gagaaagagg agaaatacta gatgagtgtg atcgctaaac
     1741 aaatgaccta caaggtttat atgtcaggca cggtcaatgg acactacttt gaggtcgaag
     1801 gcgatggaaa aggtaagccc tacgaggggg agcagacggt aaagctcact gtcaccaagg
     1861 gcggacctct gccatttgct tgggatattt tatcaccaca gtgtcagtac ggaagcatac
     1921 cattcaccaa gtaccctgaa gatatccctg actatgtaaa gcagtcattc ccggagggct
     1981 atacatggga gaggatcatg aactttgaag atggtgcagt gtgtactgtc agcaatgatt
     2041 ccagcatcca aggcaactgt ttcatctacc atgtcaagtt ctctggtttg aactttcctc
     2101 acaatggacc tgtcatgcag aagaaaacac agggctggga acccaacact gagcgtctgt
     2161 ttgcacgaga tggaatgctg ctaggaaaca actttatggc tctgaagtta gaaggaggcg
     2221 gtcactattt gtgtgaattt aaaactactt acaaggcaaa gaagcctgtg aagatgccag
     2281 ggtatcacta tgttgaccgc aaactggatg taaccaatca caacaaggat tacacttcgg
     2341 ttgagcagtg tgaaatttcc attgcacgca aacctgtggt cgcctaacaa ataaaacgaa
     2401 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     2461 gtctcagaag gtacagagac cattctcacc aataaaaaac gcccggcggc aaccgagcgt
     2521 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtctatg
     2581 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
     2641 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
     2701 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
     2761 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
     2821 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
     2881 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
     2941 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
     3001 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
     3061 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
     3121 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
     3181 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
     3241 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
     3301 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatcc caaagccttt
     3361 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
     3421 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
     3481 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
     3541 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
     3601 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
     3661 ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg
     3721 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
     3781 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
     3841 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
     3901 actttagtca
//
