LOCUS       Exported                6473 bp ds-DNA     circular SYN 10-JAN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6473)
  AUTHORS   Jonathan Pacheco
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, May 18, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6473
                     /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    2363..2411
                     /label=MCS
                     /note="MCS"
     CDS             2419..3135
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     2419..2436
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     complement(2422..2439)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     2498..2517
                     /label=J42
     primer_bind     complement(3116..3135)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     3116..3135
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     polyA_signal    3261..3382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3389..3844)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3871..3975
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3977..4334
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4185..4320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4369..5163
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5395..5442
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5771..6359
                     /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 gctagccgcc
      601 accatgccca agccaatcaa cgtccgggtg accaccatgg atgccgagct ggagtttgcc
      661 atccagccaa acaccaccgg gaagcagctc tttgatcagg tggtaaagac gattggcctc
      721 cgggaagtgt ggtacttcgg cctccagtat gtagacaata aaggatttcc tacctggctg
      781 aaacttgata aaaaggtctc cgcacaggag gtccgaaagg agaaccctgt ccagtttaaa
      841 ttccgggcca agttctaccc cgaagacgtg gccgaggaac tcatccagga catcacacag
      901 aagctcttct tcctgcaagt caaggacggg atcctcagcg acgagatcta ctgcccccca
      961 gagacagccg tgctcctggg ctcctatgcc gttcaggcca agttcggaga ttataacaag
     1021 gaaatgcaca agtctgggta cctcagctcg gagcggctga tcccccagag agtcatggac
     1081 caacacaagc tcagcaggga ccagtgggag gaccggatcc aggtgtggca cgcggaacac
     1141 cgagggatgc tcaaggacag tgctatgcta gaatacctga agattgccca ggacctggaa
     1201 atgtatggga tcaactattt cgagatcaaa aacaagaaag gaacagacct ttggcttgga
     1261 gtcgatgccc ttggacttaa catttatgag aaagatgaca agttgacccc aaagatcggc
     1321 ttcccttgga gtgagatcag gaacatctct ttcaacgaca agaagtttgt cattaagccc
     1381 atcgacaaga annnacctga ctttgtgttc tacgccccgc gcctgagaat taacaagcgg
     1441 atcctgcagc tctgcatggg gaaccatgag ctgtacatgc gccgcaggaa gcccgacacc
     1501 atcgaggtgc agcagatgaa ggcccaggct cgggaggaga agcaccagaa gcagctagag
     1561 cgacagcagt tggaaaccga gaagaagagg cgagagacgg tggagagaga aaaggagcag
     1621 atgctccggg agaaggagga gctgatgctt cggctgcagg actacgagca gaagaccaag
     1681 agggcggaga aagagctctc cgagcagatt gagaaggccc tccaactgga ggaagagagg
     1741 aggcgagccc aggaggaggc tgaacgtctg gaggccgacc gcatggccgc cctgcgggcc
     1801 aaggaagaac tcgagagaca ggcgcaggat cagataaaga gccaggagca gctggctgca
     1861 gagctggcag agtacacggc caagatcgca ctgctggagg aggcgcggag gcgcaaggag
     1921 gacgaggtag aagagtggca gcaccgggct aaagaagccc aggacgacct ggtgaagacc
     1981 aaagaggagc tgcacctggt gatgacggcc ccaccgcccc caccgccccc agtgtatgag
     2041 cctgtgaatt accacgtgca ggagggactg caggacgagg gagcagagcc tatgggctac
     2101 agtgccgagc tctccagtga gggcatcctg gatgaccgca acgaggagaa gcggatcaca
     2161 gaggcagaga agaatgagcg cgtgcagcgg cagctgctga ccctgagcaa tgagttgtcc
     2221 caggcccggg atgagaacaa gaggacccac aatgacatca tccacaacga gaacatgcgg
     2281 caaggcaggg acaagtataa gacgctgcgg caaatcaggc agggcaacac caagcaacgc
     2341 attgacgagt tcgaggccat gaagcttcga attctgcagt cgacggtacc gcgggcccgg
     2401 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2461 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2521 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2581 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2641 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2701 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2761 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2821 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2881 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2941 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3001 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3061 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3121 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     3181 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3241 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3301 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3361 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     3421 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3481 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3541 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3601 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3661 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3721 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3781 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3841 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3901 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3961 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4021 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4081 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4141 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4201 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4261 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4321 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4381 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4441 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4501 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4561 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4621 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4681 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4741 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4801 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4861 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     4921 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4981 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5041 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5101 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5161 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5221 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5281 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     5341 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5401 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5461 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5521 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5581 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5641 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5701 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5761 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5821 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5881 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5941 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6001 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6061 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6121 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6181 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6241 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6301 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6361 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6421 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
