LOCUS       Exported                5696 bp ds-DNA     circular SYN 20-OCT-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5696)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 20, 2022 from SnapGene 5.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5696
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    591..671
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      673..678
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             679..1311
                     /codon_start=1
                     /label=Homo sapiens clathrin light chain B (CLTB)
                     /translation="MADDFGFFSSSESGAPEAAEEDPAAAFLAQQESEIAGIENDEGFG
                     APAGSHAAPAQPGPTSGAGSEDMGTTVNGDVFQEANGPADGYAAIAQADRLTQEPESIR
                     KWREEQRKRLQELDAASKVTEQEWREKAKKDLEEWNQRQSEQVEKNKINNRASEEAFVK
                     ESKEETPGTEWEKVAQLCDFNPKSSKQCKDVSRLRSVLMSLKQTPLSR"
     CDS             1330..1653
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             1672..2357
                     /codon_start=1
                     /label=EGFP insert
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAA"
     polyA_signal    2484..2605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2612..3067)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3094..3198
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3200..3557
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3408..3543
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3592..4386
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4618..4665
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4994..5582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccaccgg tcgccaccat ggctgatgac tttggcttct tctcgtcgtc ggagagtggt
      721 gccccggagg cggcggagga ggacccggcg gccgccttcc tggcccagca ggagagcgag
      781 attgcaggca tagagaacga cgagggcttc ggggcacctg ccggcagcca tgcggccccc
      841 gcacagccgg gccccacgag tggggctggt tctgaggaca tggggaccac agtcaatgga
      901 gatgtgtttc aggaggccaa cggtcctgct gatggctacg cagccattgc ccaggctgac
      961 aggctgaccc aggagcctga gagcatccgc aagtggcgag aggagcagag gaaacggctg
     1021 caagagctgg atgctgcatc taaggtcacg gaacaggaat ggcgggagaa ggccaagaag
     1081 gacctggagg agtggaacca gcgccagagt gaacaagtag agaagaacaa gatcaacaac
     1141 cgggcatccg aggaggcttt cgtgaaggaa tccaaggagg agaccccagg cacagagtgg
     1201 gagaaggtgg cccagctatg tgacttcaac cccaagagca gcaagcagtg caaagatgtg
     1261 tcccgcctgc gctcggtgct catgtccctg aagcagacgc cactgtcccg cggcggatcc
     1321 ggagggtcta tgggagtgca ggtggaaacc atctccccag gagacgggcg caccttcccc
     1381 aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg aaagaaattt
     1441 gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca ggaggtgatc
     1501 cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa actgactata
     1561 tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc acatgccact
     1621 ctcgtcttcg atgtggagct tctaaaactg gaaggaggat ccggagggtc tatggtgagc
     1681 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1741 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1801 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1861 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1921 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1981 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2041 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2101 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     2161 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     2221 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2281 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2341 ttcgtgaccg ccgccggcta aagcggccgc gactctagat cataatcagc cataccacat
     2401 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2461 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2521 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2581 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     2641 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2701 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2761 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2821 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2881 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2941 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3001 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3061 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3121 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3181 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3301 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3361 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3421 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3481 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3541 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3601 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3661 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3721 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3781 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3841 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3901 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3961 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4021 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4081 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4141 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4201 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4261 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4321 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4381 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4441 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4501 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4561 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4621 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4681 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4741 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4801 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4861 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4921 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4981 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5041 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5101 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5161 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5221 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5281 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5341 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5401 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5461 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5521 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5581 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5641 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
