LOCUS       pUS252L                 3359 bp DNA     circular SYN 14-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pUS252L
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3359)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Sep 14, 2022 from SnapGene 6.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    208..229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        244..274
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     257..276
                     /label=NVC15b
     protein_bind    282..298
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(283..310)
                     /label=MVS76
     primer_bind     305..353
                     /label=MVS73
     RBS             325..330
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             338..1063
                     /codon_start=1
                     /label=fuBFP
                     /translation="MVSSGEDIFSGLVPILIELEGDVNGHRFSVRGEGYGDASNGKLEI
                     KFICTTGRLPVPWPTLVTTLSHGVQCFAKYPEHMRQNDFFKSAMPDGYVQERTISFKED
                     GTYKTRAEVKFEGEALVNRIDLKGLEFKEDGNILGHKLEYSFNSHYVYITADKNRNGLE
                     AQFRIRHNVDDGSVQLADHYQQNTPIGEGPVLLPEQHYLTTNSVLSKDPQERRDHMVLV
                     EFVTAAGLSLGMDELYKS*"
     primer_bind     complement(514..534)
                     /label=MVS69
     primer_bind     complement(520..559)
                     /label=MVS38
     primer_bind     528..547
                     /label=MVS70
                     /note="Degeneracies caused an issue for snapchat, 5' part 
                     is in fact complementary to T64"
     misc_feature    536..538
                     /label=Y66H
     primer_bind     complement(623..641)
                     /label=MVS57
     primer_bind     636..655
                     /label=MVS58
     primer_bind     complement(731..752)
                     /label=MVS59
     primer_bind     749..771
                     /label=MVS60
     primer_bind     complement(807..829)
                     /label=MVS61*
                     /note="Does not actually contain the final base, just 
                     included to get Snapchat to recognise the binding site"
     primer_bind     826..845
                     /label=MVS62
     primer_bind     complement(844..862)
                     /label=MVS63
     primer_bind     858..875
                     /label=MVS64
     primer_bind     complement(882..911)
                     /label=MVS65*
                     /note="final 5 nucleotides (lowercase) aren't in actual 
                     primer, just here to get snapchat to believe me"
     primer_bind     907..933
                     /label=MVS66*
                     /note="final 6 nucleotides (lower case) aren't in actual 
                     primer, merely a ploy to deceive snapchat"
     primer_bind     complement(926..946)
                     /label=MVS71
     primer_bind     941..959
                     /label=MVS72
     primer_bind     complement(982..1001)
                     /label=MVS67
     primer_bind     996..1015
                     /label=MVS68
     primer_bind     complement(1048..1076)
                     /label=MVS74
     primer_bind     1177..1195
                     /label=LFE7
     primer_bind     1396..1415
                     /label=LFE4
     primer_bind     1460..1476
                     /label=MVS75
     primer_bind     1460..1476
                     /label=MVS42
     promoter        1481..1509
                     /label=KanR promoter
     CDS             1613..2407
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1660..1679)
                     /label=LFE5
     primer_bind     complement(1879..1896)
                     /label=MVS49
     primer_bind     2073..2093
                     /label=LFE10
     primer_bind     2165..2173
                     /label=MVS38
     primer_bind     complement(2329..2349)
                     /label=MVS37
     primer_bind     complement(2386..2408)
                     /label=MVS43
     primer_bind     complement(2386..2407)
                     /label=LFE3
     rep_origin      2691..3279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2934..2941
                     /label=MVS73
     primer_bind     complement(3270..3283)
                     /label=NVC330
     primer_bind     join(3357..3359,1..26)
                     /label=iGEM25
ORIGIN
        1 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
       61 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
      121 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
      181 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      241 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      301 tcacggatcc gaattctaga cctcgaggag gtttcatatg gtgagcagtg gcgaagatat
      361 tttctcgggc ttggttccga ttctgatcga gctggagggc gatgtgaacg gtcatcgttt
      421 tagcgttcgc ggtgaaggtt atggcgacgc gagcaacggc aaactggaaa ttaagttcat
      481 ctgcacgacc ggtcgcctgc cggtgccttg gccgaccttg gtgacgacct tgagccatgg
      541 cgtgcagtgt tttgcgaagt atccggagca catgcgccaa aacgatttct ttaaaagtgc
      601 gatgccggac ggttacgtcc aggagcgtac catttccttc aaggaagatg gcacgtacaa
      661 aactcgcgca gaggttaagt ttgaaggtga agcgctggtc aatcgtatcg atttgaaggg
      721 tttggagttt aaagaggatg gtaacattct gggccataaa ctggagtata gcttcaacag
      781 ccattatgtt tacattacgg cagacaagaa tcgtaacggc ttggaggccc aattccgtat
      841 tcgccacaat gttgatgacg gtagcgtcca actggccgac cattaccaac agaacacccc
      901 aattggtgag ggtccggtgt tgctgccgga acaacactat ctgacgacca atagcgtgct
      961 gagcaaagat ccgcaggaac gccgtgatca catggtcctg gtggaatttg tgaccgctgc
     1021 gggcttgagc ctgggtatgg acgagctgta taagagctaa tgaaagcttg ctgcaggcat
     1081 gcaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     1141 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     1201 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg ataagctagc
     1261 ttcacgctgc cgcaagcact cagggcgcaa gggctgctaa aggaagcgga acacgtagaa
     1321 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     1381 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     1441 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     1501 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     1561 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     1621 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1681 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1741 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tccaagacga
     1801 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1861 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     1921 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     1981 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     2041 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     2101 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cggatgcccg acggcgagga
     2161 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2221 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     2281 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     2341 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     2401 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     2461 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     2521 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     2581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2821 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccc
//
