LOCUS       sequence-418158-        3910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU1-A 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  JOURNAL   unpublished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            "unpublished"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: pTU1-A_Sv4K_amilCP
FEATURES             Location/Qualifiers
     source          1..3910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(224..1018)
                     /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(335..354)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     945..964
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1250..1268
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1272..1315)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1338..1372
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1381..1392
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2065..2136
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2159..2253)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2354..3304)
                     /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      3352..3574
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ctgctatgtg gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca
       61 gtctgaccac ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa
      121 ggcagcggta tcatcaacaa gcaaagaaaa aaggcgacag attactctgt cgcctttttt
      181 cctgactcat aaccaagcca attctcgaac cccagagtcc cgctcagaag aactcgtcaa
      241 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
      301 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
      361 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
      421 tttccaccat gatattcggc aagcaggcat cgccgtgggt cacgacgaga tcctcgccgt
      481 cgggcatacg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgatctt
      541 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
      601 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
      661 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
      721 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
      781 cagccgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
      841 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcga ccctgcgctg
      901 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
      961 atagcctctc cacccaagcc gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     1021 gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg
     1081 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
     1141 cagctggcaa ttccggcacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     1201 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     1261 cacgaggcag aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttccgtct
     1321 caatctctat agagaccttt acagctagct cagtcctagg tattatgcta gctactagag
     1381 aaagaggaga aatactagat gagtgtgatc gctaaacaaa tgacctacaa ggtttatatg
     1441 tcaggcacgg tcaatggaca ctactttgag gtcgaaggcg atggaaaagg taagccctac
     1501 gagggggagc agacggtaaa gctcactgtc accaagggcg gacctctgcc atttgcttgg
     1561 gatattttat caccacagtg tcagtacgga agcataccat tcaccaagta ccctgaagat
     1621 atccctgact atgtaaagca gtcattcccg gagggctata catgggagag gatcatgaac
     1681 tttgaagatg gtgcagtgtg tactgtcagc aatgattcca gcatccaagg caactgtttc
     1741 atctaccatg tcaagttctc tggtttgaac tttcctccca atggacctgt catgcagaag
     1801 aaaacacagg gctgggaacc caacactgag cgtctgtttg cacgagatgg aatgctgcta
     1861 ggaaacaact ttatggctct gaagttagaa ggaggcggtc actatttgtg tgaatttaaa
     1921 actacttaca aggcaaagaa gcctgtgaag atgccagggt atcactatgt tgaccgcaaa
     1981 ctggatgtaa ccaatcacaa caaggattac acttcggttg agcagtgtga aatttccatt
     2041 gcacgcaaac ctgtggtcgc ctaacaaata aaacgaaagg atcagtcgaa agactgggcc
     2101 tttcgtttta tctgttgttt gtcggtgaac gctctcggtc tcatgtttgc cagagacgat
     2161 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     2221 ctgaggtcat tactggatct atcaacagna gtctatggac agttttccct ttgatatgta
     2281 acggtgaaca gttgttctac ttttgtttgt tagtcttgat gcttcactga tagatacaag
     2341 agccataaga acctcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt
     2401 gtttttgcgt gagccatgag aacgaaccat tgagatcata cttactttgc atgtcactca
     2461 aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt
     2521 ttcttagtcc gttatgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat
     2581 tcatttttat ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa
     2641 cttggaaaat caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc
     2701 tgtaagtgtt taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact
     2761 catggtagtt attttcaagc attaacatga acttaaattc atcaaggcta atctctatat
     2821 ttgccttgtg agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag
     2881 agtatttgtt ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact
     2941 ggaaaagata aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact
     3001 tggcatagtt tgtccactgg aaaatcccaa agcctttaac caaaggattc ctgatttcca
     3061 cagttctcgt catcagctct ctggttgctt tagctaatac accataagca ttttccctac
     3121 tgatgttcat catctgagcg tattggttat aagtgaacga taccgtccgt tctttccttg
     3181 tagggttttc aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat
     3241 gctccgttaa gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag
     3301 acatacatct caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc
     3361 aatgataatt actagtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct
     3421 ttgctggaaa acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg
     3481 ttttttttgt ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga
     3541 taaaaagaat agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt
     3601 acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag
     3661 gcttaagtag caccctcgca agctcggttg cggccgcaat cgggcaaatc gctgaatatt
     3721 ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc
     3781 tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc ttttatggat
     3841 tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct cagggcgttt
     3901 tatggcgggt
//
