LOCUS       sequence-387108-        7305 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 7305)
  AUTHORS   Edie S, Zaghloul NA, Leitch CC, Klinedinst DK, Lebron J, Thole JF, 
            McCallion AS, Katsanis N, Reeves RH
  TITLE     Survey of Human Chromosome 21 Gene Expression Effects on Early 
            Development in Danio rerio.
  JOURNAL   G3 (Bethesda). 2018 Jul 2;8(7):2215-2223. doi: 
            10.1534/g3.118.200144.
  PUBMED    29760202
REFERENCE   2  (bases 1 to 7305)
  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"; doi: "10.1534/g3.118"; 
            journalName: "G3 (Bethesda)"; date: "2018-07-2- 2"; volume: "8"; 
            issue: "7"; pages: "2215-2223"
COMMENT     Sequence Label: COL6A1_pCSdest
FEATURES             Location/Qualifiers
     source          1..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    4..28
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3152..3176)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3220..3354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3244..3263)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3298..3317
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3467..3487)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3467..3485)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3506..3522)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3506..3522)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3519..3541)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3530..3546
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3554..3560,3561..3578,3579..3584))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3737..3754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3908..4496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3988..4007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4671..5462,5463..5531))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5294..5313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5532..5636)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5662..6117)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5799..5820)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6011..6030
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6188..7172
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        7207..7225
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7207..7224
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 atgccaactt tgtacaaaaa agcaggctcc gcggccgccc ccttcaccat gagggcggcc
       61 cgtgctctgc tgcccctgct gctgcaggcc tgctggacag ccgcgcagga tgagccggag
      121 accccgaggg ccgtggcctt ccaggactgc cccgtggacc tgttctttgt gctggacacc
      181 tctgagagcg tggccctgag gctgaagccc tacggggccc tcgtggacaa agtcaagtcc
      241 ttcaccaagc gcttcatcga caacctgagg gacaggtact accgctgtga ccgaaacctg
      301 gtgtggaacg caggcgcgct gcactacagt gacgaggtgg agatcatcca aggcctcacg
      361 cgcatgcctg gcggccgcga cgcactcaaa agcagcgtgg acgcggtcaa gtactttggg
      421 aagggcacct acaccgactg cgctatcaag aaggggctgg agcagctcct cgtggggggc
      481 tcccacctga aggagaataa gtacctgatt gtggtgaccg acgggcaccc cctggagggc
      541 tacaaggaac cctgtggggg gctggaggat gctgtgaacg aggccaagca cctgggcgtc
      601 aaagtcttct cggtggccat cacacccgac cacctggagc cgcgtctgag catcatcgcc
      661 acggaccaca cgtaccggcg caacttcacg gcggctgact ggggccagag ccgcgacgca
      721 gaggaggcca tcagccagac catcgacacc atcgtggaca tgatcaaaaa taacgtggag
      781 caagtgtgct gctccttcga atgccagcct gcaagaggac ctccggggct ccggggcgac
      841 cccggctttg agggagaacg aggcaagccg gggctcccag gagagaaggg agaagccgga
      901 gatcctggaa gacccgggga cctcggacct gttgggtacc agggaatgaa gggagaaaaa
      961 gggagccgtg gggagaaggg ctccagggga cccaagggct acaagggaga gaagggcaag
     1021 cgtggcatcg acggggtgga cggcgtgaag ggggagatgg ggtacccagg cctgccaggc
     1081 tgcaagggct cgcccgggtt tgacggcatt caaggacccc ctggccccaa gggagacccc
     1141 ggcgcctttg gactgaaagg agaaaagggc gagcctggag ctgacgggga ggcggggaga
     1201 ccagggagct cgggaccatc tggagacgag ggccagccgg gagagcctgg gccccccgga
     1261 gagaaaggag aggcgggcga cgaggggaac ccaggacctg acggtgcccc cggggagcgg
     1321 ggtggccctg gagagagagg accacggggg accccaggca cacggggacc aagaggagac
     1381 cctggtgaag ctggcccgca gggtgatcag ggaagagaag gccccgttgg tgtccctgga
     1441 gacccgggcg aggctggccc tatcggacct aaaggctacc gaggcgatga gggtccccca
     1501 gggtccgagg gtgccagagg agccccagga cctgccggac cccctggaga cccggggctg
     1561 atgggtgaaa ggggagaaga cggccccgct ggaaatggca ccgagggctt ccccggcttc
     1621 cccgggtatc cgggcaacag gggcgctccc gggataaacg gcacgaaggg ctaccccggc
     1681 ctcaaggggg acgagggaga agccggggac cccggagacg ataacaacga cattgcaccc
     1741 cgaggagtca aaggagcaaa ggggtaccgg ggtcccgagg gcccccaggg acccccagga
     1801 caccaaggac cgcctgggcc ggacgaatgc gagattttgg acatcatcat gaaaatgtgc
     1861 tcttgctgtg aatgcaagtg cggccccatc gacctcctgt tcgtgctgga cagctcagag
     1921 agcattggcc tgcagaactt cgagattgcc aaggacttcg tcgtcaaggt catcgaccgg
     1981 ctgagccggg acgagctggt caagttcgag ccagggcagt cgtacgcggg tgtggtgcag
     2041 tacagccaca gccagatgca ggagcacgtg agcctgcgca gccccagcat ccggaacgtg
     2101 caggagctca aggaagccat caagagcctg cagtggatgg cgggcggcac cttcacgggg
     2161 gaggccctgc agtacacgcg ggaccagctg ctgccgccca gcccgaacaa ccgcatcgcc
     2221 ctggtcatca ctgacgggcg ctcagacact cagagggaca ccacaccgct caacgtgctc
     2281 tgcagccccg gcatccaggt ggtctccgtg ggcatcaaag acgtgtttga cttcatccca
     2341 ggctcagacc agctcaatgt catttcttgc caaggcctgg caccatccca gggccggccc
     2401 ggcctctcgc tggtcaagga gaactatgca gagctgctgg aggatgcctt cctgaagaat
     2461 gtcaccgccc agatctgcat agacaagaag tgtccagatt acacctgccc catcacgttc
     2521 tcctccccgg ctgacatcac catcctgctg gacggctccg ccagcgtggg cagccacaac
     2581 tttgacacca ccaagcgctt cgccaagcgc ctggccgagc gcttcctcac agcgggcagg
     2641 acggaccccg cccacgacgt gcgggtggcg gtggtgcagt acagcggcac gggccagcag
     2701 cgcccagagc gggcgtcgct gcagttcctg cagaactaca cggccctggc cagtgccgtc
     2761 gatgccatgg actttatcaa cgacgccacc gacgtcaacg atgccctggg ctatgtgacc
     2821 cgcttctacc gcgaggcctc gtccggcgct gccaagaaga ggctgctgct cttctcagat
     2881 ggcaactcgc agggcgccac gcccgctgcc atcgagaagg ccgtgcagga agcccagcgg
     2941 gcaggcatcg agatcttcgt ggtggtcgtg ggccgccagg tgaatgagcc ccacatccgc
     3001 gtcctggtca ccggcaagac ggccgagtac gacgtggcct acggcgagag ccacctgttc
     3061 cgtgtcccca gctaccaggc cctgctccgc ggtgtcttcc accagacagt ctccaggaag
     3121 gtggcgctgg gctagaaggg tgggcgcgcc gacccagctt tcttgtacaa agtggggatc
     3181 ctctcgagcc tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac
     3241 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3301 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3361 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc
     3421 gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt
     3481 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3541 tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat
     3601 gagtgagcta actcayatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3661 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3721 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3781 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3841 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgacgcgtaa ctcacgttaa
     4561 gggattttgg tcatgagctt gcgccgtccc gtcaagtcag mgttatgctt aaattaaaaa
     4621 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4681 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4741 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4801 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4861 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4921 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4981 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5041 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5101 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5161 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5221 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5281 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5341 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5401 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5461 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5521 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5581 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5641 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5701 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5761 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5821 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5881 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5941 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     6001 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     6061 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     6121 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6181 tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt
     6241 ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt
     6301 ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc
     6361 ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag
     6421 gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc
     6481 catattggct atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat
     6541 atggctattg gccaggttca atactatgta ttggccctat gccatatagt attccatata
     6601 tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg
     6661 gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg
     6721 tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga
     6781 cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc
     6841 accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt
     6901 gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt
     6961 ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc
     7021 aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag
     7081 gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg
     7141 aggtctatat aagcaatgct cgtttaggga accgccattc tgcctgggga cgtcggagca
     7201 agcttgattt aggtgacact atagaataca agctacttgt tctttttgca ggatcccata
     7261 gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttata
//
