LOCUS       Exported                6599 bp ds-DNA     circular SYN 14-OCT-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6599)
  AUTHORS   Oliver
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Oct 17, 2022 from SnapGene 5.1.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6599
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      607..616
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             613..1329
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1:  613 ..  615 / #05fd14
                        2:  616 ..  618 / #05fd14 / 1a
                        3:  619 .. 1329 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1231..1233
                     /label=A206K
                     /note="color: #a6acb3"
     CDS             1366..3219
                     /codon_start=1
                     /label=hNET
                     /note="color: #a6acb3"
                     /translation="MLLARMNPQVQPENNGADTGPEQPLRARKTAELLVVKERNGVQCL
                     LAPRDGDAQPRETWGKKIDFLLSVVGFAVDLANVWRFPYLCYKNGGGAFLIPYTLFLII
                     AGMPLFYMELALGQYNREGAATVWKICPFFKGVGYAVILIALYVGFYYNVIIAWSLYYL
                     FSSFTLNLPWTDCGHTWNSPNCTDPKLLNGSVLGNHTKYSKYKFTPAAEFYERGVLHLH
                     ESSGIHDIGLPQWQLLLCLMVVVIVLYFSLWKGVKTSGKVVWITATLPYFVLFVLLVHG
                     VTLPGASNGINAYLHIDFYRLKEATVWIDAATQIFFSLGAGFGVLIAFASYNKFDNNCY
                     RDALLTSSINCITSFVSGFAIFSILGYMAHEHKVNIEDVATEGAGLVFILYPEAISTLS
                     GSTFWAVVFFVMLLALGLDSSMGGMEAVITGLADDFQVLKRHRKLFTFGVTFSTFLLPL
                     FCITKGGIYVLTLLDTFAAGTSILFAVLMEAIGVSWFYGVDRFSNDIQQMMGFRPGLYW
                     RLCWKFVSPAFLLFVVVVSIINFKPLTYDDYIFPPWANWVGWGIALSSMVLVPIYVIYK
                     FLSTQGSLWERLAYGITPENEHHLVAQRDIRQFQLQHWLAI"
     misc_difference 2734..2736
                     /label=A457P
                     /note="color: #a6acb3; direction: RIGHT"
     polyA_signal    3387..3508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(3515..3970)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3997..4101
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        4103..4460
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      4311..4446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4495..5289
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5521..5568
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     rep_origin      5897..6485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc aaactgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attccatgct tctggcgcgg
     1381 atgaacccgc aggtgcagcc cgagaacaac ggggcggaca cgggtccaga gcagcccctt
     1441 cgggcgcgca aaactgcgga gctgctggtg gtgaaggagc gcaacggcgt ccagtgcctg
     1501 ctggcgcccc gcgacggcga cgcgcagccc cgggagacct ggggcaagaa gatcgacttc
     1561 ctgctgtccg tagtcggctt cgcagtggac ctggccaacg tgtggcgctt cccctacctc
     1621 tgctacaaga acggcggcgg tgccttcttg atcccgtaca cactgttcct tatcatcgcg
     1681 gggatgcccc tgttctacat ggagctggct ctgggacagt acaaccggga gggggctgcc
     1741 accgtttgga agatctgccc attcttcaaa ggcgttggct atgctgtcat cctgatcgcc
     1801 ctgtacgttg gcttctacta caacgtcatc atcgcctggt cactctacta cctcttctcc
     1861 tccttcaccc tcaacctgcc ctggaccgac tgtggccaca cctggaacag ccccaactgt
     1921 accgacccca agctcctcaa tggctccgtg cttggcaacc acaccaagta ctccaagtac
     1981 aagttcacgc cggcagccga gttttatgag cgtggtgtcc tgcaccttca cgagagcagc
     2041 gggattcatg acatcggcct gccccagtgg cagctcttgc tctgtctgat ggtcgtcgtc
     2101 atcgtcttgt attttagcct ctggaaaggg gtgaagacat caggaaaggt ggtgtggatc
     2161 acagccacgc tgccttactt cgtgctgttc gtgctcctgg tccatggggt caccctgccc
     2221 ggagcctcca atggcatcaa tgcctacctg cacatcgact tctaccgctt gaaagaggcc
     2281 acggtatgga ttgatgccgc aactcagata tttttttcct tgggggctgg atttggagta
     2341 ttgattgcat ttgccagtta caacaaattt gacaacaact gttacaggga tgccctgctg
     2401 accagcagca tcaactgtat caccagcttc gtctctgggt tcgccatctt ctccatcctt
     2461 ggttacatgg cccatgaaca caaggtcaac attgaggatg tggccacaga aggagctggc
     2521 ctagtgttca tcctgtatcc agaggccatt tctaccctgt ctggatctac attctgggct
     2581 gttgtgtttt tcgtcatgct cctggcgctg ggccttgaca gctcaatggg aggcatggag
     2641 gctgtcatca cgggcctggc cgatgacttc caggtcctga agcgacaccg gaaactcttc
     2701 acatttggcg tcaccttcag cactttcctt ctccccctgt tctgcataac caagggtgga
     2761 atttacgtct tgaccctcct ggacaccttt gctgcgggca cctccatcct ttttgctgtc
     2821 ctcatggaag ccatcggagt ttcctggttt tatggagtgg acaggttcag caacgacatc
     2881 cagcagatga tggggttcag gccgggtcta tactggagac tgtgctggaa gttcgtcagt
     2941 cctgccttcc tcctgttcgt ggttgtggtc agcatcatca acttcaagcc actcacctac
     3001 gacgactaca tcttcccgcc ctgggccaac tgggtggggt ggggcatcgc cctgtcctcc
     3061 atggtcctgg tgcccatcta cgtcatctat aagttcctca gcacgcaggg ctctctttgg
     3121 gagagactgg cctatggcat cacgccagag aacgagcacc acctggtggc tcagagggac
     3181 atcagacagt tccagttgca acactggctg gccatctgag cctgcctgga ggagaaggag
     3241 gaacctctag agggcccgga tccaccggat ctagataact gatcataatc agccatacca
     3301 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3361 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3421 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3481 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     3541 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3601 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3661 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3721 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3781 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3841 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3901 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3961 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4021 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4081 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4141 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4201 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4261 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4321 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4381 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4441 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4501 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4561 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4621 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4681 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4741 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4801 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4861 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4921 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4981 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5041 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5101 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5161 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5221 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5281 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5341 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5401 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5461 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5521 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5581 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5641 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5701 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5761 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5821 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5881 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5941 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6001 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6061 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6121 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6181 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6241 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6301 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6361 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6421 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6481 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6541 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
