LOCUS       sequence-418183-        3910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-b backbone with kanamycin resistance and pSC101-var5 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  JOURNAL   unpublished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            "unpublished"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_Sv5K_amilCP
FEATURES             Location/Qualifiers
     source          1..3910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(557..1351)
                     /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(668..687)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1278..1297
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1583..1601
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1605..1648)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1671..1705
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1714..1725
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2398..2469
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2492..2586)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2687..3637)
                     /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      3685..3907
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
       61 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg ttgcggccgc
      121 aatcgggcaa atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt
      181 ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg
      241 cactacaggc gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc
      301 gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat ctgacttttt
      361 gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat tatcccgtga
      421 caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa caagcaaaga
      481 aaaaaggcga cagattactc tgtcgccttt tttcctgact cataaccaag ccaattctcg
      541 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
      601 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
      661 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
      721 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
      781 catcgccgtg ggtcacgacg agatcctcgc cgtcgggcat acgcgccttg agcctggcga
      841 acagttcggc tggcgcgagc ccctgatgat cttcgtccag atcatcctga tcgacaagac
      901 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
      961 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     1021 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     1081 agtcccttcc cgcttcagtg acaacgtcga gcacagccgc gcaaggaacg cccgtcgtgg
     1141 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     1201 tcttgacaaa aagaaccggg cgaccctgcg ctgacagccg gaacacggcg gcatcagagc
     1261 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gccgccggag
     1321 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     1381 cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     1441 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggc accccttgta
     1501 ttactgttta tgtaagcaga cagttttatt gttgccacct gacgtctaag aaaccattat
     1561 tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa
     1621 atccttagct ttcgctaagg atgatttcgg tctcactata tctagagacg tttacagcta
     1681 gctcagtcct aggtattatg ctagctacta gagaaagagg agaaatacta gatgagtgtg
     1741 atcgctaaac aaatgaccta caaggtttat atgtcaggca cggtcaatgg acactacttt
     1801 taggtcgaag gcgatggaaa aggtaagccc tacgaggggg agcagacggt aaagctcact
     1861 gtcaccaagg gcggacctct gccatttgct tgggatattt tatcaccaca gtgtcagtac
     1921 ggaagcatac cattcaccaa gtaccctgaa gatatccctg actatgtaaa gcagtcattc
     1981 ccggagggct atacatggga gaggatcatg aactttgaag atggtgcagt gtgtactgtc
     2041 agcaatgatt ccagcatcca aggcaactgt ttcatctacc atgtcaagtt ctctggtttg
     2101 aactttcctc ccaatggacc tgtcatgcag aagaaaacac agggctggga acccaacact
     2161 gagcgtctgt ttgcacgaga tggaatgctg ctaggaaaca actttatggc tctgaagtta
     2221 gaaggaggcg gtcactattt gtgtgaattt aaaactactt acaaggcaaa gaagcctgtg
     2281 aagatgccag ggtatcacta tgttgaccgc aaactggatg taaccaatca caacaaggat
     2341 tacacttcgg ttgagcagtg tgaaatttcc attgcacgca aacctgtggt cgcctaacaa
     2401 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2461 aacgctctcc gtctcagaag gtacagagac cattctcacc aataaaaaac gcccggcggc
     2521 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     2581 ggagtctatg gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt
     2641 tgttagtctt gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt
     2701 atttagccag tatgttctct agtgtgattc gttgtttttg cgtgagccat gagaacgaac
     2761 cattgagatc atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct
     2821 gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat
     2881 ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt
     2941 cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc
     3001 ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg
     3061 gtttcaaaac ccattggtta agccttttaa actcatggta gttattttca agcattaaca
     3121 tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta
     3181 gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat
     3241 gttccagatt atattttatg aattttttta actggaaaag ataaggcaat atctcttcac
     3301 taaaaactaa ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct
     3361 caaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg
     3421 ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt
     3481 tataagtgaa cgataccgtc cgttctttcc atgtagggtt ttcaatcgtg gggttgagta
     3541 gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg
     3601 ctagttcatt tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat
     3661 tttaatcact ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt
     3721 gagttgtggg tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta
     3781 gaccctctgt aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta
     3841 taatttatag aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt
     3901 ataactcact
//
