LOCUS       Exported                4817 bp ds-DNA     circular SYN 20-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4817)
  AUTHORS   joeyr
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Sep 29, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4817
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..580
                     /label=ER-RA F Primer
                     /note="Forward primer to amplify ERM from ERM-APEX2."
     primer_bind     complement(545..562)
                     /label=ER-RA R Vector Primer
                     /note="Reverse primer to amplify Pex3-GA backbone."
     regulatory      557..566
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             563..643
                     /codon_start=1
                     /label=ERM
                     /translation="MDPVVVLGLCLSCLLLLSLWKQSYGGG"
     primer_bind     complement(626..696)
                     /label=ER-RA R Primer
                     /note="Reverse primer to amplify ERM from ERM-APEX2."
     CDS             644..676
                     /codon_start=1
                     /label=Linker
                     /translation="GGGGSGGGGSA"
     CDS             677..1369
                     /codon_start=1
                     /label=GA
                     /translation="MVSKSEEVIKEFMRFKVRLEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPLIMYGSKMYVKHPADVPDYMKLSFPEGFKWERVMHFEDGG
                     LVTATQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTLGWDYATERLYPEEGVLKGELL
                     GRLKLKDGGLNLVESKTIYMAKKPVQLPGYYFVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLGHGTGST"
     primer_bind     677..698
                     /label=ER-RA F Vector Primer
                     /note="Reverse primer to amplify Pex3-GA backbone."
     polyA_signal    1560..1681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1688..2143)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2170..2274
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2276..2633
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2484..2619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2668..3462
                     /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    3694..3741
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4070..4658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgtgccgcca ccatggaccc tgtggtcgtg ctggggctgt gcctgagctg
      601 tctgctgctg ctgagcctgt ggaagcagtc ctacggaggc gggggtggtg gtggtagtgg
      661 tggtggtggt agtgctatgg tgagcaagag cgaggaggtc atcaaagagt tcatgcgctt
      721 caaggtgcgc ttggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
      781 gggccgccct tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct
      841 gcccttcgcc tgggacatcc tgtcccccct gatcatgtac ggctccaaga tgtacgtgaa
      901 gcaccccgcc gacgtccccg attacatgaa gctgtccttc cccgagggct tcaagtggga
      961 gcgcgtgatg cacttcgagg acggcggtct ggtgaccgcg acacaggact cctccctgca
     1021 ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc ccgacggccc
     1081 cgtaatgcag aagaagacct tgggctggga ttatgccacc gagcgcctgt accccgaaga
     1141 aggcgtgctg aagggcgagc tcctggggcg cctgaagctg aaggacggcg gcctcaacct
     1201 ggtggagtcc aagaccatct acatggccaa gaagcccgtg caactgcccg gctactactt
     1261 cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     1321 cgagcgctcc gagggccgcc accacctggg gcatggcacc ggcagcacct aatagaagct
     1381 tggtaaaccg ctgatcagcc tcgactgcag tcgacggtac cgcgggcccg ggatccaccg
     1441 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     1501 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1561 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1621 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1681 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1741 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     1801 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     1861 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     1921 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     1981 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2041 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2101 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     2161 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2221 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     2281 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2341 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2401 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2461 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2521 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     2581 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     2641 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     2701 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     2761 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     2821 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     2881 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     2941 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3001 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3061 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3121 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3181 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3241 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3301 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3361 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     3421 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     3481 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     3541 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3601 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     3661 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     3721 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     3781 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     3841 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     3901 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3961 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4021 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4081 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4141 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4201 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4261 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4321 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4381 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4441 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4501 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4561 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4621 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4681 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4741 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
     4801 tcattagttc atagccc
//
