LOCUS       Exported                8228 bp ds-DNA     circular SYN 07-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8228)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Aug 8, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8228
                     /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"
     CDS             616..642
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             709..4884
                     /codon_start=1
                     /label=CLIP-170
                     /translation="MSMLKPSGLKAPTKILKPGSTALKTPTAVVAPVEKTISSEKASST
                     PSSETQEEFVDDFRVGERVWVNGNKPGFIQFLGETQFAPGQWAGIVLDEPIGKNDGSVA
                     GVRYFQCEPLKGIFTRPSKLTRKVQAEDEANGLQTTPASRATSPLCTSTASMVSSSPST
                     PSNIPQKPSQPAAKEPSATPPISNLTKTASESISNLSEAGSIKKGERELKIGDRVLVGG
                     TKAGVVRFLGETDFAKGEWCGVELDEPLGKNDGAVAGTRYFQCQPKYGLFAPVHKVTKI
                     GFPSTTPAKAKANAVRRVMATTSASLKRSPSASSLSSMSSVASSVSSRPSRTGLLTETS
                     SRYARKISGTTALQEALKEKQQHIEQLLAERDLERAEVAKATSHVGEIEQELALARDGH
                     DQHVLELEAKMDQLRTMVEAADREKVELLNQLEEEKRKVEDLQFRVEEESITKGDLEVA
                     TVSEKSRIMELEKDLALRVQEVAELRRRLESNKPAGDVDMSLSLLQEISSLQEKLEVTR
                     TDHQREITSLKEHFGAREETHQKEIKALYTATEKLSKENESLKSKLEHANKENSDVIAL
                     WKSKLETAIASHQQAMEELKVSFSKGLGTETAEFAELKTQIEKMRLDYQHEIENLQNQQ
                     DSERAAHAKEMEALRAKLMKVIKEKENSLEAIRSKLDKAEDQHLVEMEDTLNKLQEAEI
                     KVKELEVLQAKCNEQTKVIDNFTSQLKATEEKLLDLDALRKASSEGKSEMKKLRQQLEA
                     AEKQIKHLEIEKNAESSKASSITRELQGRELKLTNLQENLSEVSQVKETLEKELQILKE
                     KFAEASEEAVSVQRSMQETVNKLHQKEEQFNMLSSDLEKLRENLADMEAKFREKDEREE
                     QLIKAKEKLENDIAEIMKMSGDNSSQLTKMNDELRLKERDVEELQLKLTKANENASFLQ
                     KSIEDMTVKAEQSQQEAAKKHEEEKKELERKLSDLEKKMETSHNQCQELKARYERATSE
                     TKTKHEEILQNLQKTLLDTEDKLKGAREENSGLLQELEELRKQADKAKAAQTAEDAMQI
                     MEQMTKEKTETLASLEDTKQTNAKLQNELDTLKENNLKNVEELNKSKELLTVENQKMEE
                     FRKEIETLKQAAAQKSQQLSALQEENVKLAEELGRSRDEVTSHQKLEEERSVLNNQLLE
                     MKKRESKFIKDADEEKASLQKSISITSALLTEKDAELEKLRNEVTVLRGENASAKSLHS
                     VVQTLESDKVKLELKVKNLELQLKENKRQLSSSSGNTDTQADEDERAQESQIDFLNSVI
                     VDLQRKNQDLKMKVEMMSEAALNGNGDDLNNYDSDDQEKQSKKKPRLFCDICDCFDLHD
                     TEDCPTQAQMSEDPPHSTHHGSRGEERPYCEICEMFGHWATNCNDDETF"
     polyA_signal    5016..5137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5144..5599)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5626..5730
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5732..6089
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5940..6075
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6124..6918
                     /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    7150..7197
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7526..8114
                     /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 ccggtcgcca ccatgtaccc atacgatgtt ccagattacg cttccggact cagatctcga
      661 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccat gagtatgcta
      721 aagccaagtg ggcttaaggc ccccaccaag atcctgaagc ctggaagcac agctctgaag
      781 acacctacgg ctgttgtagc tccagtagaa aaaaccatat ccagtgaaaa agcatcaagc
      841 actccatcat ctgagactca ggaggaattt gtggatgact ttcgagttgg ggagcgagtt
      901 tgggtgaatg gaaataagcc tggatttatc cagtttcttg gagaaaccca gtttgcacca
      961 ggccagtggg ctggaattgt tttagatgaa cccataggca agaacgatgg ttcggtggca
     1021 ggagttcggt atttccagtg tgaaccttta aagggcatat ttacccgacc ttcaaagtta
     1081 acaaggaagg tgcaagcaga agatgaagct aatggcctgc agacaacgcc cgcctcccga
     1141 gctacttcac cgctgtgcac ttctacggcc agcatggtgt cttcctcccc ctccacccct
     1201 tcaaacatcc ctcagaaacc atcacagcca gcagcaaagg aaccttcagc tacgcctccg
     1261 atcagcaacc ttacaaaaac tgccagtgaa tctatctcca acctttcaga ggctggctca
     1321 atcaagaaag gagaaagaga gctcaaaatc ggagacagag tattggttgg tggcactaag
     1381 gctggtgtag tccggtttct tggggagacc gactttgcca agggggagtg gtgtggcgtg
     1441 gagttagatg agccacttgg gaagaatgat ggcgctgttg ctggaacaag gtattttcag
     1501 tgtcaaccca aatatggctt gttcgctcct gtccacaaag ttaccaagat tggcttccct
     1561 tccactacac cagccaaagc caaggccaac gcagtgaggc gagtgatggc gaccacgtcc
     1621 gccagcctga agcgcagccc ttctgcctct tccctcagct ccatgagctc agtggcctcc
     1681 tctgtgagca gcaggcccag tcggacagga ctattgactg aaacctcctc ccgttacgcc
     1741 aggaagatct ccggtaccac tgccctccag gaggccctga aggagaagca gcagcacatt
     1801 gagcagctgc tggcggaacg ggatctggag agggcggagg tggccaaggc cacgagccac
     1861 gtgggggaga tagagcagga gctagctctg gcccgggacg gacatgacca gcatgtcctg
     1921 gaattggaag ccaaaatgga ccagctgcga acaatggtgg aagctgctga cagggagaag
     1981 gtggagcttc tcaaccagct tgaagaggag aaaaggaagg ttgaggacct tcagttccgg
     2041 gttgaagaag aatcaattac caaaggtgat cttgaggtgg ctacagtttc agaaaagtca
     2101 cgtataatgg aactggagaa agacctagca ttgagagtac aggaagtagc tgagctccga
     2161 agaaggctag agtccaataa gcctgctggg gatgtggaca tgtcactttc ccttttgcaa
     2221 gagataagct ctttgcaaga aaagttagaa gtcacccgta ctgaccacca gagagaaata
     2281 acttctctga aggagcattt tggagcccgg gaagaaactc atcagaagga gataaaggct
     2341 ctgtataccg ccacggaaaa gctttccaaa gagaacgagt cattgaaaag caagctggag
     2401 catgccaaca aagagaactc agatgtgata gctctatgga agtccaaact ggagactgcc
     2461 atcgcatccc accagcaggc gatggaagaa ctgaaggtat ctttcagcaa agggcttgga
     2521 acagagacgg cagaatttgc tgaactaaaa acacaaatag agaaaatgag actagattac
     2581 caacacgaaa tagaaaattt gcagaatcaa caagactctg aacgggctgc ccatgctaaa
     2641 gagatggaag ccttgagggc taaactgatg aaagttatta aagaaaagga aaacagtctg
     2701 gaagccatca ggtcgaaact ggacaaagca gaagaccagc atctcgtaga aatggaagac
     2761 acgttaaaca aattacagga agctgaaata aaggtaaagg agctagaggt actgcaagcc
     2821 aaatgcaatg aacaaaccaa ggttattgat aattttacat cacagctcaa ggctactgaa
     2881 gaaaagctct tggatcttga tgcacttcgg aaagccagtt ccgaaggtaa atcggaaatg
     2941 aagaaactta gacagcagct tgaggcagct gagaaacaga ttaaacattt agagattgaa
     3001 aagaatgctg aaagtagcaa ggctagtagc attaccagag agctccaggg gagagagcta
     3061 aagcttacta accttcagga aaatttgagt gaagtcagtc aagtgaaaga gactttggaa
     3121 aaagaacttc agattttgaa agaaaagttt gctgaagctt cagaggaggc agtctctgtt
     3181 cagagaagta tgcaagaaac tgtaaataag ttacaccaaa aggaggaaca gtttaacatg
     3241 ctgtcttctg acttggagaa gctgagagaa aacttagcag atatggaggc aaaatttaga
     3301 gagaaagatg agagagaaga gcagctgata aaggcaaagg aaaaactgga aaatgacatt
     3361 gcagaaataa tgaagatgtc aggagataac tcttctcagc tgacaaaaat gaacgatgaa
     3421 ttacgtctga aagaaagaga tgtagaagaa ttacagctaa aacttacaaa ggctaatgaa
     3481 aatgcaagtt ttctgcaaaa aagtattgag gacatgactg tcaaagctga acagagccag
     3541 caagaagcag ctaaaaagca tgaggaagaa aagaaagaat tggagaggaa attgtcggac
     3601 ctggaaaaga aaatggaaac aagccacaac cagtgtcagg agctgaaagc caggtatgag
     3661 agagccactt ctgagacaaa aaccaagcat gaagaaatcc tacagaacct ccagaagacg
     3721 ctgctggaca cagaggacaa gctgaagggc gcacgggagg agaacagtgg cttgctgcag
     3781 gagctggagg agctgagaaa gcaagccgac aaagccaaag ctgctcaaac agcggaagat
     3841 gccatgcaga taatggaaca gatgaccaaa gagaagactg agactctggc ctccttggag
     3901 gacaccaagc aaacaaatgc aaaactacag aatgaattgg acacacttaa agaaaacaac
     3961 ttgaaaaatg tggaagagct gaacaaatca aaagaactcc tgactgtaga gaatcaaaaa
     4021 atggaagaat ttaggaaaga aatagaaacc ctaaagcagg cagcagctca gaagtcccag
     4081 cagctttcag cgttgcaaga agagaacgtt aaacttgctg aggagctggg gagaagcagg
     4141 gacgaagtca caagtcatca aaagctggaa gaagaaagat ctgtgctcaa taatcagttg
     4201 ttagaaatga aaaaaagaga atccaagttc ataaaagacg cagatgaaga gaaagcttcc
     4261 ttgcagaaat ccatcagtat aactagtgcc ttactcacag aaaaggatgc cgagctggag
     4321 aaactgagaa atgaggtcac agtgctcagg ggagaaaacg cctctgccaa gtccttgcat
     4381 tcagttgttc agactctaga gtctgataag gtgaagctcg agctcaaggt aaagaacttg
     4441 gagcttcaac tcaaagaaaa caagaggcag ctcagcagct cctcaggtaa tacagacact
     4501 caggcagacg aggatgaaag agcccaggag agtcagattg atttcctaaa ttcagtaata
     4561 gtggaccttc aaaggaagaa tcaagacctc aagatgaagg tggagatgat gtcagaagca
     4621 gccctgaatg ggaacgggga tgacctaaac aattatgaca gtgatgatca ggagaaacag
     4681 tccaagaaga aacctcgcct cttctgtgac atttgtgact gctttgatct ccacgacaca
     4741 gaggattgtc ctacccaggc acagatgtca gaggaccctc cccattccac acaccatggc
     4801 agtcggggtg aggaacgccc atactgtgaa atctgtgaga tgtttggaca ctgggccacc
     4861 aactgcaatg acgacgaaac cttctaggat ccaccggatc tagataactg atcataatca
     4921 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4981 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     5041 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5101 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     5161 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5221 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5281 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5341 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5401 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5461 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5521 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5581 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5641 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5701 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     5761 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5821 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5881 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5941 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6001 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6061 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     6121 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     6181 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6241 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6301 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6361 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6421 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6481 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6541 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6601 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     6661 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6721 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6781 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6841 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6901 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6961 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     7021 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     7081 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     7141 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     7201 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     7261 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     7321 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     7381 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7441 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7501 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7561 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7621 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     7681 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7741 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7801 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7861 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7921 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7981 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8041 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8101 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8161 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8221 ccatgcat
//
