LOCUS       Exported                4783 bp ds-DNA     circular SYN 29-MAR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4783)
  AUTHORS   Hammond Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4783
                     /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"
     primer_bind     519..539
                     /label=CMV-F
     misc_feature    591..614
                     /label=MCS
                     /note="MCS"
     CDS             618..650
                     /codon_start=1
                     /label=Lyn11
                     /note="Lyn11"
                     /translation="MGSIKSKGKDS"
     CDS             651..665
                     /codon_start=1
                     /label=GGSGG
                     /translation="GGSGG"
     misc_feature    666..695
                     /label=Helical linker
     CDS             696..704
                     /codon_start=1
                     /label=GGSGG
                     /translation="GGS"
     CDS             705..719
                     /codon_start=1
                     /label=GGSGG
                     /translation="GGSGG"
     primer_bind     complement(715..731)
                     /label=Lyn-R
     misc_feature    720..721
                     /label=MCS
                     /note="MCS"
     primer_bind     729..746
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     misc_feature    732..1445
                     /label=EGFP
                     /note="EGFP"
     primer_bind     complement(732..749)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     732..749
                     /label=FPi-F2.0
     primer_bind     1377..1397
                     /label=A41
     primer_bind     complement(1426..1445)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     1426..1445
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     complement(1477..1496)
                     /label=SV40-pArev
     primer_bind     1530..1552
                     /label=A42
     polyA_signal    1571..1692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1699..2154)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2181..2285
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2287..2644
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2679..3473
                     /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"
     terminator      3648..3917
                     /label=TK_PA
                     /note="TK_polyadenylation_signal"
     misc_feature    4065..4684
                     /label=Ori_pBR322
                     /note="pBR322_replication_origin"
     rep_origin      4081..4669
                     /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 atctaccatg ggatctataa aatcaaaagg gaaagacagc ggtggctctg
      661 gaggggaagc cgctgctaga gaagctgccg caagaggcgg atcagggggt tccggaggcc
      721 tcgccgccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      781 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      841 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      901 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      961 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1021 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1081 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1141 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1201 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1261 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1321 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1381 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1441 acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     1501 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     1561 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1621 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1681 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     1741 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     1801 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     1861 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     1921 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     1981 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2041 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2101 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2161 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2221 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2281 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2341 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2401 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2461 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2521 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2581 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2641 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2701 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2761 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2821 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     2881 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2941 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3001 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3061 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3121 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3181 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     3241 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3301 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3361 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3421 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3481 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3541 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3601 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     3661 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     3721 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3781 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3841 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3901 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     3961 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4021 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4081 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4141 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4201 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4261 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4321 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4381 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4441 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4501 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4561 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4621 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4681 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4741 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
