LOCUS       pNCM39_(pEGFP-C1        7083 bp DNA     circular SYN 26-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   nichd_dir_cssc_sub
  TITLE     Direct Submission
  JOURNAL   Exported May 26, 2024 from SnapGene Viewer 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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..636
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(646..648,649..1359)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             1378..3516
                     /label=WT POLH CDS
     CDS             3517..3738
                     /label=Delta GG NEDD8 CDS
     polyA_signal    3871..3992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3999..4454)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4481..4585
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4587..4944
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4587..4778
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      4795..4930
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4979..5773
                     /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    6005..6052
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6381..6969
                     /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 ccatgccaaa gaagaagcga aaggtagcag atccagtgag caagggcgag
      661 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      721 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      781 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      841 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      901 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      961 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1021 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1081 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1141 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1201 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1261 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1321 gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagtg
     1381 gctactggac aggatcgagt ggttgctctc gtggacatgg actgtttttt tgttcaagtg
     1441 gagcagcggc aaaatcctca tttgaggaat aaaccttgtg cagttgtaca gtacaaatca
     1501 tggaagggtg gtggaataat tgcagtgagt tatgaagctc gtgcatttgg agtcactaga
     1561 agtatgtggg cagatgatgc taagaagtta tgtccagatc ttctactggc acaagttcgt
     1621 gagtcccgtg ggaaagctaa cctcaccaag taccgggaag ccagtgttga agtgatggag
     1681 ataatgtctc gttttgctgt gattgaacgt gccagcattg atgaggctta cgtagatctg
     1741 accagtgctg tacaagagag actacaaaag ctacaaggtc agcctatctc ggcagacttg
     1801 ttgccaagca cttacattga agggttgccc caaggcccta caacggcaga agagactgtt
     1861 cagaaagagg ggatgcgaaa acaaggctta tttcaatggc tcgattctct tcagattgat
     1921 aacctcacct ctccagacct gcagctcacc gtgggagcag tgattgtgga ggaaatgaga
     1981 gcagccatag agagggagac tggttttcag tgttcagctg gaatttcaca caataaggtc
     2041 ctggcaaaac tggcctgtgg actaaacaag cccaaccgcc aaaccctggt ttcacatggg
     2101 tcagtcccac agctcttcag ccaaatgccc attcgcaaaa tccgtagtct tggaggaaag
     2161 ctaggggcct ctgtcattga gatcctaggg atagaataca tgggtgaact gacccagttc
     2221 actgaatccc agctccagag tcattttggg gagaagaatg ggtcttggct atatgccatg
     2281 tgccgaggga ttgaacatga tccagttaaa cccaggcaac tacccaaaac cattggctgt
     2341 agtaagaact tcccaggaaa aacagctctt gctactcggg aacaggtaca atggtggctg
     2401 ttgcaattag cccaggaact agaggagaga ctgactaaag accgaaatga taatgacagg
     2461 gtagccaccc agctggttgt gagcattcgt gtacaaggag acaaacgcct cagcagcctg
     2521 cgccgctgct gtgcccttac ccgctatgat gctcacaaga tgagccatga tgcatttact
     2581 gtcatcaaga actgtaatac ttctggaatc cagacagaat ggtctcctcc tctcacaatg
     2641 cttttcctct gtgctacaaa attttctgcc tctgcccctt catcttctac agacatcacc
     2701 agcttcttga gcagtgaccc aagttctctg ccaaaggtgc cagttaccag ctcagaagct
     2761 aagacccagg gaagtggccc agcggtgaca gccactaaga aagcaaccac gtctctggaa
     2821 tcattcttcc aaaaagctgc agaaaggcag aaagttaaag aagcttcgct ttcatctctt
     2881 actgctccca ctcaggctcc catgagcaat tcaccatcca agccctcatt accttttcaa
     2941 accagtcaaa gtacaggaac tgagcccttc tttaagcaga aaagtctgct tctaaagcag
     3001 aaacagctta ataattcttc agtttcttcc ccccaacaaa acccatggtc caactgtaaa
     3061 gcattaccaa actctttacc aacagagtat ccagggtgtg tccctgtttg tgaaggggtg
     3121 tcgaagctag aagaatcctc taaagcaact cctgcagaga tggatttggc ccacaacagc
     3181 caaagcatgc acgcctcttc agcttccaaa tctgtgctgg aggtgactca gaaagcaacc
     3241 ccaaatccaa gtcttctagc tgctgaggac caagtgccct gtgagaagtg tggctccctg
     3301 gtaccggtat gggatatgcc agaacacatg gactatcatt ttgcattgga gttgcagaaa
     3361 tcctttttgc agccccactc ttcaaacccc caggttgttt ctgccgtatc tcatcaaggc
     3421 aaaagaaatc ccaagagccc tttggcctgc actaataaac gccccaggcc tgagggcatg
     3481 caaacattgg aatcattttt taagccatta acacatatgc taattaaagt gaagacgctg
     3541 accggaaagg agattgagat tgacattgaa cctacagaca aggtggagcg aatcaaggag
     3601 cgtgtggagg agaaagaggg aatcccccca caacagcaga ggctcatcta cagtggcaag
     3661 cagatgaatg atgagaagac agcagctgat tacaagattt taggtggttc agtccttcac
     3721 ctggtgttgg ctctgagata aggatccacc ggatctagat aactgatcat aatcagccat
     3781 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3841 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3901 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3961 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     4021 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4081 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4141 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4201 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4261 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4321 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4381 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4441 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4501 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4561 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4621 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4681 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4741 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4801 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4861 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4921 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4981 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5041 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5101 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5161 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5221 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5281 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5341 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5401 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5461 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5521 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5581 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5641 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5701 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5761 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5821 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5881 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5941 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6001 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6061 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6121 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6181 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6241 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6301 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6361 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6421 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6481 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     6541 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6601 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6661 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6721 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6781 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6841 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6901 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6961 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7021 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     7081 cat
//
