LOCUS       sequence-418180-        3910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU2-b backbone with kanamycin resistance and pSC101-var7 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  JOURNAL   unpublished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            "unpublished"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_Sv7K_amilCP
FEATURES             Location/Qualifiers
     source          1..3910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(543..1337)
                     /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(654..673)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1264..1283
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1569..1587
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1591..1634)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1657..1691
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1700..1711
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2384..2455
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2478..2572)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2673..3623)
                     /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      3671..3893
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa
       61 accctaaagg cttaagtagc accctcgcaa gctcggttgc ggccgcaatc gggcaaatcg
      121 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
      181 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct
      241 tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc
      301 agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt
      361 cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac
      421 tggctaatgc acccagtaag gcagcggtat catcaacaag caaagaaaaa aggcgacaga
      481 ttactctgtc gccttttttc ctgactcata accaagccaa ttctcgaacc ccagagtccc
      541 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
      601 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
      661 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
      721 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccgtgggtc
      781 acgacgagat cctcgccgtc gggcatacgc gccttgagcc tggcgaacag ttcggctggc
      841 gcgagcccct gatgatcttc gtccagatca tcctgatcga caagaccggc ttccatccga
      901 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
      961 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     1021 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     1081 tcagtgacaa cgtcgagcac agccgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     1141 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     1201 accgggcgac cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     1261 tgtgcccagt catagccgaa tagcctctcc acccaagccg ccggagaacc tgcgtgcaat
     1321 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
     1381 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     1441 accttaccag agggcgcccc agctggcaat tccggcaccc cttgtattac tgtttatgta
     1501 agcagacagt tttattgttg ccacctgacg tctaagaaac cattattatc atgacattaa
     1561 cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg
     1621 ctaaggatga tttcggtctc actatatcta gagacgttta cagctagctc agtcctaggt
     1681 attatgctag ctactagaga aagaggagaa atactagatg agtgtgatcg ctaaacaaat
     1741 gacctacaag gtttatatgt caggcacggt caatggacac tactttgagg tcgaaggcga
     1801 tggaaaaggt aagccctacg agggggagca gacggtaaag ctcactgtca ccaagggcgg
     1861 acctctgcca tttgcttggg atattttatc accacagtgt cagtacggaa gcataccatt
     1921 caccaagtac cctgaagata tccctgacta tgtaaagcag tcattcccgg agggctatac
     1981 atgggagagg atcatgaact ttgaagatgg tgcagtgtgt actgtcagca atgattccag
     2041 catccaaggc aactgtttca tctaccatgt caagttctct ggtttgaact ttcctcccaa
     2101 tggacctgtc atgcagaaga aaacacaggg ctgggaaccc aacactgagc gtctgtttgc
     2161 acgagatgga atgctgctag gaaacaactt tatggctctg aagttagaag gaggcggtca
     2221 ctatttgtgt gaatttaaaa ctacttacaa ggcaaagaag cctgtgaaga tgccagggta
     2281 tcactatgtt gaccgcaaac tggatgtaac caatcacaac aaggattaca cttcggttga
     2341 gcagtgtgaa atttccattg cacgcaaacc tgtggtcgcc taacaaataa aacgaaagga
     2401 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctccgtct
     2461 cagaaggtac agagaccatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
     2521 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tctatggaca
     2581 gttttccctt tgatatgtaa cggtgaacag ttgttctact tttgtttgtt agtcttgatg
     2641 cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt agccagtatg
     2701 ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatac
     2761 ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt
     2821 aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt aggaatctga tgtaatggtt
     2881 gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc
     2941 atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca
     3001 accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat
     3061 tggttaagcc ttttaaactc atggtagtta tttccaagca ttaacatgaa cttaaattca
     3121 tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac
     3181 cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat
     3241 tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa aactaattct
     3301 aatttttcgc ttgagaactt ggcatagtct gtccactgga aaatctcaaa gcctttaacc
     3361 aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca
     3421 ccataagcat tttccctact gatgttcatc atctgagcgt attggttata agtgaacgat
     3481 accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat
     3541 aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct
     3601 ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac
     3661 caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt tgtgggtatc
     3721 tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat
     3781 tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata
     3841 aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt
     3901 tagtcagttc
//
