LOCUS       sequence-339891-        5008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5008)
  TITLE     TA Rapoport plasmids
REFERENCE   2  (bases 1 to 5008)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAc-GFPC1-Sec61beta
FEATURES             Location/Qualifiers
     source          1..5008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        104..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     562..582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      650..659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             656..1372
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     CDS             1388..1678
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
                     RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     polyA_signal    1839..1960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1876..1895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1930..1949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1967..2422)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2104..2125)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2316..2335
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2449..2553
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2555..2912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2555..2746
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2578..2598)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2763..2898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2947..3741
                     /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"
     CDS             2947..3741
                     /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"
     primer_bind     complement(3001..3020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3611..3630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3929..3948)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3973..4020
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4349..4937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4838..4857
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      601 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
      661 gagcaagggc gccgagctgt tcaccggcat cgtgcccatc ctgatcgagc tgaatggcga
      721 tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag ggcgatgcca cctacggcaa
      781 gctgaccctg aagttcatct gcaccaccgg caagctgcct gtgccctggc ccaccctggt
      841 gaccaccctg agctacggcg tgcagtgctt ctcacgctac cccgatcaca tgaagcagca
      901 cgacttcttc aagagcgcca tgcctgaggg ctacatccag gagcgcacca tcttcttcga
      961 ggatgacggc aactacaagt cgcgcgccga ggtgaagttc gagggcgata ccctggtgaa
     1021 tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc aacatcctgg gcaataagat
     1081 ggagtacaac tacaacgccc acaatgtgta catcatgacc gacaaggcca agaatggcat
     1141 caaggtgaac ttcaagatcc gccacaacat cgaggatggc agcgtgcagc tggccgacca
     1201 ctaccagcag aataccccca tcggcgatgg ccctgtgctg ctgcccgata accactacct
     1261 gtccacccag agcgccctgt ccaaggaccc caacgagaag cgcgatcaca tgatctactt
     1321 cggcttcgtg accgccgccg ccatcaccca cggcatggat gagctgtaca agtccggact
     1381 cagatctatg cctggtccga cccccagtgg cactaacgtg ggatcctcag ggcgctctcc
     1441 cagcaaagca gtggccgccc gggcggcggg atccactgtc cggcagagga aaaatgccag
     1501 ctgtgggaca aggagtgcag gccgcacaac ctcggcaggc accgggggga tgtggcgatt
     1561 ctacacagaa gattcacctg ggctcaaagt tggccctgtt ccagtattgg ttatgagtct
     1621 tctgttcatc gcttctgtat ttatgttgca catttggggc aagtacactc gttcgtagga
     1681 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     1741 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1801 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1861 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1921 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     1981 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2041 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2101 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2161 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2221 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2281 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2341 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2401 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2461 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2521 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2581 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2641 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2701 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2761 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2821 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2881 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     2941 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3001 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3061 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3121 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3181 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3241 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3301 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3361 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3421 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3481 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3541 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3601 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3661 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3721 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3781 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3841 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3901 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3961 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4021 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4081 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4141 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4201 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4261 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4321 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4381 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4441 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4501 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4561 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4621 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4681 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4741 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4801 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4861 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4921 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4981 ggccttttgc tcacatgttc tttcctgc
//
