LOCUS       pNCM36_(pEGFP-C1        4761 bp DNA     circular SYN 24-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4761)
  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..4761
                     /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"
     misc_feature    616..645
                     /label=Insert
                     /note="added to pEGFP-C1"
     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"
     misc_feature    1360..1425
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1549..1670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1677..2132)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2159..2263
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2265..2622
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2265..2456
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      2473..2608
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2657..3451
                     /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    3683..3730
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4059..4647
                     /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 atctcgagct
     1381 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa
     1441 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     1501 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     1561 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1621 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     1681 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1741 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     1801 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1861 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     1921 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     1981 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2041 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2101 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     2161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     2281 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2341 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2401 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2461 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2521 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2581 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     2641 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2701 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2761 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2821 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2881 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2941 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3001 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3061 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3121 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3181 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3241 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3301 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3361 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3421 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3481 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3541 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3601 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     3661 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     3721 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3781 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3841 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     3901 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3961 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4021 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4081 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4141 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4201 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4261 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4321 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4381 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4441 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4501 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4561 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4621 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4681 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4741 aaccgtatta ccgccatgca t
//
