LOCUS       sequence-418155-        3909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type TU1-A 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 3909)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3909)
  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            "unpublished"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_Sv7K_amilCP
FEATURES             Location/Qualifiers
     source          1..3909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(560..1354)
                     /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(671..690)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1281..1300
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1586..1604
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1608..1651)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        1674..1707
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             1716..1727
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2400..2471
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2494..2588)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2689..3639)
                     /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      3687..3909
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
       61 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cggttgcggc
      121 cgcaatcggg caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc
      181 tgtctttttc gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa
      241 tggcactaca ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag
      301 cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc tatctgactt
      361 tttgctgttc agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg
      421 tgacaggtca ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaagcaa
      481 agaaaaaagg cgacagatta ctctgtcgcc ttttttcctg actcataacc aagccaattc
      541 tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
      601 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
      661 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
      721 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
      781 aggcatcgcc gtgggtcacg acgagatcct cgccgtcggg catacgcgcc ttgagcctgg
      841 cgaacagttc ggctggcgcg agcccctgat gatcttcgtc cagatcatcc tgatcgacaa
      901 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
      961 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     1021 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     1081 gccagtccct tcccgcttca gtgacaacgt cgagcacagc cgcgcaagga acgcccgtcg
     1141 tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt
     1201 cggtcttgac aaaaagaacc gggcgaccct gcgctgacag ccggaacacg gcggcatcag
     1261 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagccgccg
     1321 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     1381 gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     1441 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggcacccctt
     1501 gtattactgt ttatgtaagc agacagtttt attgttgcca cctgacgtct aagaaaccat
     1561 tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa
     1621 aaaatcctta gctttcgcta aggatgattt ccgtctcaat ctctatagag acctttacag
     1681 ctagctcagt cctagtatta tgctagctac tagagaaaga ggagaaatac tagatgagtg
     1741 tgatcgctaa acaaatgacc tacaaggttt atatgtcagg cacggtcaat ggacactact
     1801 ttgaggtcga aggcgatgga aaaggtaagc cctacgaggg ggagcagacg gtaaagctca
     1861 ctgtcaccaa gggcggacct ctgccatttg cttgggatat tttatcacca cagtgtcagt
     1921 acggaagcat accattcacc aagtaccctg aagatatccc tgactatgta aagcagtcat
     1981 tcccggaggg ctatacatgg gagaggatca tgaactttga agatggtgca gtgtgtactg
     2041 tcagcaatga ttccagcatc caaggcaact gtttcatcta ccatgtcaag ttctctggtt
     2101 tgaactttcc tcccaatgga cctgtcatgc agaagaaaac acagggctgg gaacccaaca
     2161 ctgagcgtct gtttgcacga gatggaatgc tgctaggaaa caactttatg gctctgaagt
     2221 tagaaggagg cggtcactat ttgtgtgaat ttaaaactac ttacaaggca aagaagcctg
     2281 tgaagatgcc agggtatcac tatgttgacc gcaaactgga tgtaaccaat cacaacaagg
     2341 attacacttc ggttgagcag tgtgaaattt ccattgcacg caaacctgtg gtcgcctaac
     2401 aaataaaacg aaaggatcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     2461 tgaacgctct cggtctcatg tttgccagag acgattctca ccaataaaaa acgcccggcg
     2521 gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa
     2581 caggagtcta tggacagttt tccctttgat atgtaacggt gaacagttgt tctacttttg
     2641 tttgttagtc ttgatgcttc actgatagat acaagagcca taagaacctc agatccttcc
     2701 gtatttagcc agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga
     2761 accattgaga tcatacttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag
     2821 ctgaattttt gcagttaaag catcgtgtag tgtttttctt agtccgttat gtaggtagga
     2881 atctgatgta atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag
     2941 ttcggttacg agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg
     3001 gcggcctcgc ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat
     3061 tggtttcaaa acccattggt taagcctttt aaactcatgg tagttatttc caagcattaa
     3121 catgaactta aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt
     3181 tagttctttt aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac
     3241 atgttccaga ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc
     3301 actaaaaact aattctaatt tttcgcttga gaacttggca tagtctgtcc actggaaaat
     3361 ctcaaagcct ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt
     3421 tgctttagct aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg
     3481 gttataagtg aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag
     3541 tagtgccaca cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat
     3601 cgctagttca tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg
     3661 attttaatca ctataccaat tgagatgggc tagtcaatga taattactag tccttttcct
     3721 ttgagttgtg ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc
     3781 tagaccctct gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt
     3841 tataatttat agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt
     3901 gtataactc
//
