LOCUS       Exported                5354 bp ds-DNA     circular SYN 20-OCT-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  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..5354
                     /organism="synthetic DNA construct"
                     /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..1296
                     /codon_start=1
                     /label=Homo sapiens clathrin light chain B (CLTB)
                     /translation="MADDFGFFSSSESGAPEAAEEDPAAAFLAQQESEIAGIENDEGFG
                     APAGSHAAPAQPGPTSGAGSEDMGTTVNGDVFQEANGPADGYAAIAQADRLTQEPESIR
                     KWREEQRKRLQELDAASKVTEQEWREKAKKDLEEWNQRQSEQVEKNKINNRASEEAFVK
                     ESKEETPGTEWEKVAQLCDFNPKSSKQCKDVSRLRSVLMSLKQ"
     CDS             1315..2019
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="EELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICT
                     TGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKT
                     RAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKI
                     RHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTA
                     AGITLGMDELYK"
     polyA_signal    2142..2263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2270..2725)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2752..2856
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2858..3215
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3066..3201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3250..4044
                     /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    4276..4323
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4652..5240
                     /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 aagcaggggg gtggcggcag ctccgaggag
     1321 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1381 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1441 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1501 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1561 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1621 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1681 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1741 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1801 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1861 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     1921 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1981 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     2041 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2101 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2161 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2221 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     2281 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2341 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2401 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2461 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2521 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2581 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2641 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     2701 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2761 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2821 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     2881 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2941 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3061 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3121 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3181 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3241 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3301 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3361 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3421 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3481 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3541 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3601 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3661 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3721 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     3781 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3841 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3901 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3961 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4021 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4081 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4141 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4201 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4261 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4321 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4381 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4441 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     4501 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4561 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4621 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4681 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4741 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4801 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4861 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4921 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4981 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5041 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5101 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5161 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5221 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5281 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5341 ttaccgccat gcat
//
